Barbe-Nicole Clicquot, née Ponsardin, was widowed at 27. At the end of the 18th century, according to her status, she was supposed to marry a second time or grieve until her death, embroidering pillows and receiving guests. She did not feel the need - her father was a wealthy gentleman, and only one daughter remained from her marriage to Francois Clicquot. But the widow Clicquot loved winemaking and believed that she could fulfill her husband's dream and build a famous wine house. With this confidence, she was able to infect her own father and father-in-law, who lent money to her enterprise, former employees and companions of her husband, as well as Emperor Napoleon and Tsar Alexander the First. The history of her business is a series of failures and struggles with circumstances, weather, laws, sanctions and prejudice. "The Secret" read the book by Tilar Mazzeo and tells how the most famous champagne in the world came to be.
How did champagne come about?
The history of champagne, like any romanticized product, is full of lies and omissions. For example, Pierre Perignon, a Benedictine monk, did not invent it at all. The legend that for the first time sparkling wine began to be produced in 1660 in the abbey of Oville appeared at the end of the 19th century - it was invented by marketers at the Moёt house in order to better sell wine under the Dom Pérignon brand. The legend arose not entirely from scratch, indeed, in the cellars of the abbey, wine with bubbles sometimes ripened in cold winters, but the house of Pérignon did not approve of this and tried to get rid of the sparkling. And they loved wine with bubbles at that time not in France, but in England - by 1660 there already existed a small market for the production and sale of sparkling wine.
Another deception is that real champagne, which was adored by kings and courtiers, is not at all dry. It was about 4-5 times sweeter than the sweetest modern wine. In addition, it was mostly dark pink in color. The book of the historian Tilar Mazzeo about the widow Clicquot is filled with such revelations, because there was catastrophically little information about the widow herself. Her household and accounting books were perfectly preserved, but neither personal diaries nor love letters were found, so her fate had to be restored bit by bit.
Enterprise start
Barbe-Nicole was born in Reims in 1777, 7 years after the marriage of King Louis XVI and Marie Antoinette. The Queen of France, who adored fun, also adored wine with bubbles, fortified and more intoxicating than usual. At that time, Champagne, where Barbe-Nicole was from, already produced several thousand bottles of sparkling wine a year for the royal court and the nobility. The production of wine at that time was carried out by small households that marked their barrels with a brand, and their bottles with colorful sealing wax, just to distinguish them from those made by their neighbors.
Barbe-Nicole's father, a wealthy entrepreneur who was engaged in the textile industry, dreamed of intermarrying with the nobility, but the revolution forced him to declare himself a Jacobin and an opponent of the monarchy. He married off his eldest daughter, Barbe-Nicole, to the son of another wealthy manufacturer from Reims, Mr. Clicquot. The Clicquot family was also engaged in textiles, but also traded in wine - they bought barrels from producers in Champagne and resold them to other regions of France and a little abroad. At that time, this was a standard business scheme - it was important for winegrowers to sell wine faster in order to free up warehouses for new barrels and not be responsible for a damaged product. At that time, wine was practically not bottled, all bottles were made by hand, they were very fragile and of different sizes. In addition, until 1720, Reims was not allowed by law to bottle wine.
Francois Clicquot, the young husband of Barbe-Nicole, decided to reform the family wine business. Firstly, he was going to start exporting, and secondly, he decided that his company should also do the cultivation of grapes and wine production. As a wedding gift, she and Barbe-Nicole received solid land, including vineyards. In 1801, François decided that his own wine should account for a quarter of the total sales. In addition, he planned to produce expensive bottled wine - bottled wine could be sold for three times more than the exact same wine in barrels.
To find clients at the end of the 18th and beginning of the 19th centuries, one had to go to them with their own wine and conduct a tasting - shake for a month in a carriage, spend the night in dubious inns, suffer from cold, insects, a long road, separation from family. While François and his sales agent, Louis Bonat, tried to convince important gentlemen in Europe to order their wine, Barbe-Nicole raised the baby and oversaw sales at the local market. In the morning she went to visit the vineyards - to get good wine, you need to harvest at dawn, when the berries are wet and heavy from the dew. It was her favorite time of the day, so she wrote in letters to her husband.
The Clicquots decided to grow and process their own grapes after Napoleon, having already become emperor, visited Champagne, stayed at his father Barbe-Nicole's hotel and announced that he was going to develop French winemaking. It is known that Napoleon patronized the Moёt family, which produced, among other things, several tens of thousands of bottles of sparkling wine per year. The emperor appreciated bubbles and believed that the wine business could improve the country's economy, which was shaken after the revolution and the next round of the Anglo-French war, which ended in 1802. To spur the market, Napoleon commissioned Jean-Antoine Chaptal, a chemist and French Interior Minister, a treatise on wine production.
The Art of Making, Keeping and Improving Wine is considered a classic work on winemaking and the advice from it is still used today. For the beginning of the 19th century, this was a revolutionary work that regulated and standardized many processes that had not been recorded anywhere before and were transmitted from father to son. The Clicquots, of course, had a copy of the treatise, and they carefully followed all the recommendations. By the way, Barbe-Nicole showed a talent for blending wine - she was an excellent taster and distinguished very subtle shades of taste, which made it possible to compose exquisite bouquets from different grape varieties grown in different conditions.
In general, the fact that Barbe-Nicole was in business with her husband was an exception to the rule. If under Louis XVI there were examples of women entrepreneurs and wives involved in the management of family companies, then under Napoleon ladies, especially noble and wealthy ones, were ordered to engage in appropriate pious things - charity, needlework, education of children, balls and receptions. The woman was supposed to be a nameless decoration of the world, if someone's name was heard - this caused rumors. The only exception was for widows. Widows, on the one hand, had the respect and social status of married women, on the other hand, they received the right to conduct business like men. There were quite a few widows in the wine industry who grew grapes, made wine and sold it to distributors. Moreover, many of them were engaged in sparkling - the market segment was so small, and the production was so risky that men from large wine houses simply did not want to spend their energy on it.
Francois for several years of work experienced, it seems, all the hardships on himself. He arranged for deliveries to Great Britain - there, during the war, French wine was banned, and the British yearned for it, so they made generous orders. But in 1803 the war began again, and trade with England again became difficult. The summer of 1802 was very hot, 80% of the champagne bottles burst in the cellars, unable to withstand the heat. Louis, a sales agent who worked with the Clicquot family, went to Germany and then to Russia, counting on good sales, but was deceived in his expectations - the Germans and Russians ordered much less wine than he expected. All this bored François. He was generally prone to melancholy, although between attacks he was an energetic and cheerful person. Probably modern psychologists would have diagnosed him with bipolar disorder. In 1805 he caught typhus and died in agony. Barb-Nicole became his sole heir.
In a way, she was lucky, neither she nor François had brothers. Perhaps if there were other young men in the family, Barb-Nicole would not even think about taking matters into her own hands and would simply marry a second time, as her father wanted. A month after the death of François, Louis rushed to Reims (at that time he got very quickly, almost at lightning speed from St. Petersburg) and began to persuade the widow Clicquot not to leave the enterprise. He finally managed to make the necessary contacts in Russia, and he hoped to convert them into good sales. So the widow Clicquot began to turn into a brand.
Another attempt
Father-in-law, old Clicquot, supported his daughter-in-law and introduced her to an old friend, businessman Alexander Jerome Forno. Together they and Barbe-Nicole invested 80,000 francs in the venture. In terms of today's dollars, this is about $ 2 million. In those days, the worker received about 400 francs a year ($ 8,000), the seller - about 20,000 a year. Clicquot invested three-quarters of this amount, her partner invested the rest - mainly in wine and capital goods. Now the company sold practically only its own wine, taking only a quarter from neighboring farmers for sale.
In 1806, the business climate in France was quite harsh. In 1803, the Napoleonic Wars began, and since 1805 the emperor opposed the coalition, which included England, Russia, Sweden and Naples. Many roads were blocked, countries imposed sanctions against each other, the rules of the game changed frequently, as neighbors supported one side or the other. Under these conditions, Clicquot collected orders for 55,000 bottles of champagne ($3 million at today's exchange rate). Their partners decided to carry them through Amsterdam - Holland was a neutral country, ships left the port in Amsterdam to all parts of Europe and to Russia.
Together with the cargo, Louis Bon went, and luck let him down - not only days, but hours were not enough for the laden ships to sail. The port of Amsterdam was closed due to martial law, French ships were not released, and the wine was stuck in a warehouse. Hopes that the blockade would soon be lifted were quickly fading. The summer was hot again, and Clicquot's merchandise went bad. The widow suffered enormous losses, not only could her wine not be sold, she had to pay for an expensive warehouse and keep a chartered ship under sail in case the blockade fell. It was possible to send the cargo by smuggling on an English or American ship, but there was a risk of losing the entire batch - champagne in such a large amount could only be transported from France, and the emperor forbade trading with enemies.
The situation was complicated by the fact that the market itself was collapsing. The war pumped money out of Europe - no one was interested in champagne anymore, there was no time for luxury. Bon reached Russia and, thanks to his connections, found out that Empress Elizaveta Alekseevna was in a position - there was hope that the heir to the throne would finally be born, and on this occasion a feast would be arranged, it would not be possible to do without sparkling wine. But alas, a girl was born, who soon died - it did not work out to sell the wine. Bon, meanwhile, was accused of spying for Napoleon and almost exiled to Siberia, he constantly warned his employer in letters that she should not touch politics and write only about business, because his freedom and life were under threat. It was only in 1808 that 50,000 bottles were delivered to St. Petersburg.
When a fragile peace was established between France and Russia, where the widow Clicquot was already well known and managed to fall in love with her wine, nature let her down again - 1809 turned out to be a poor harvest, and in 1810 the partner left Barbe-Nicole. She thought about liquidating the enterprise, but this time her father persuaded her not to give up. He credited his daughter, who was now left alone not only in family life, but also in business, and Clicquot managed to buy 10,000 bottles, 125,000 corks and six dozen casks for 30,000 francs. She became an independent noble woman who single-handedly managed a fairly large international business - an outstanding example for the Napoleonic era.
The first thing the widow did when she became the sole head of her own firm was to put things in order in the accounts and ledgers. Secondly, it refocused on the local market, reduced the share of expensive but unreliable champagne and increased the share of simple but high-quality homemade table wines. By December 1810, when the widow Clicquot was 33, she had practically paid off the debts of the previous joint venture, regained her old suppliers and customers, sold a fair amount of her good wine to the French for New Year's celebrations. Meeting the year 1811, she was ready for it to bring her good luck. And he brought.
Happy Star
Pierre Bezukhov observed a comet, "which, as they said, foreshadowed all sorts of horrors and the end of the world," while driving through Moscow at night. She appeared above Prechistenka, and Pierre "joyfully, with tears in his eyes, looked at this bright star." Champagne, produced in 1811, is found in "Eugene Onegin", in the novel by Valentin Pikul "To each his own" and in other works. This is, of course, the wine "Veuve Clicquot". Barbe-Nicole was finally lucky - the summer turned out to be not hot, the wine and champagne of 1811 turned out to be incredibly transparent, with a honey aftertaste. The Big Comet flying over the vineyards added mystery to it (and added several tens of francs to the price). Clicquot considered the bottles of this vintage to be the pearls of her production.
Unfortunately, the political situation again did not allow hope to sell them at the price they deserved. In fact, in 1812 Clicquot managed to sell only 80% of the amount of wine that her unfortunate husband could sell in 1805. But then he fell ill and died from what he experienced because of a commercial failure.
By the end of 1813, the people of Reims found the war at their doorstep. Clicquot was in despair - her cellars were overflowing with wine that could not be sold. She waited in horror for the angry, hungry military - no matter from which side - to enter the city and loot her vaults. This will mean complete ruin. Most of all, she feared for the same champagne of 1811, which was almost ripe and would have brought a fabulous income in peacetime. When the Russians finally arrived, it turned out that they were not at all brutalized hordes of savages. Prince Sergei Alexandrovich Volkonsky, major of the Arkhangelsk regiment, was appointed commandant of Reims, he forbade robberies and strictly followed order. When he left the city after the armistice, the city authorities presented him with a casket studded with diamonds, in gratitude for wisdom and justice.
The officers of the Russian army did not take away, they bought wine from Barb-Nicole. Yes, many did it on credit, but the widow Clicquot willingly lent them bottles. She saw it as an investment - soon they would return home and order expensive champagne from her for holidays and anniversaries. The irony was that all these years, the widow Clicquot was chasing buyers and hated the war, which did not allow her to trade at full strength. And now the war itself brought an army of clients to it. By the way, her rival Jean-Remy Moet also understood how important it was to treat the military: “These officers, ruining me today, will bring me a fortune tomorrow,” he wrote in his diary.
light streak
The comet seemed to bring long-awaited luck to the widow Clicquot - all adventures began to work out for her. In the spring of 1814, she decided to smuggle a batch of the best champagne to Königsberg, where the Russian nobility celebrated the tsar's birthday. The bottles arrived intact, and they were sold out at the port for an inflated price. Her ships were no longer shipwrecked, buyers found her themselves and demanded more and more wine. Russia became the main market for several years, but it was in Europe that the widow was called Grande Dame - the Great Lady. From 1790 to 1830, sales of champagne in the world grew by 1000% - from several hundred thousand bottles a year to 5 million. And the merit of Barbe-Nicole in this was colossal. Widow Clicquot was the only woman manufacturer with a huge production volume and a wide trading network.
The widow came up with the idea of serving champagne in narrow tall glasses - they used to drink it from flattened bowls - and this dish soon became fashionable around the world. She was the first of the producers to start gluing bright labels on bottles so that buyers could distinguish her wine immediately and unmistakably - it was she who chose the signature orange color. Finally, Barbe-Nicole invented a method of getting rid of sediment - riddling, which is still used today. It was Clicquot who came up with a cabinet in which all bottles of ripening champagne should stand neck down at different angles and a procedure according to which they must be periodically removed, turned over and inserted into other nests at a new angle.
Clicquot was an exceptional woman, a talented businesswoman and inventor, but she was not a feminist. Clicquot fully shared the point of view of society on women and believed that it would be better for them to be married, she did not allow her daughter to do business and bequeathed the whole business to Eduard Werle, the manager whom she hired when she began to grow old and weaken. She believed that she had achieved all the successes by force - if it were not for the death of her husband and the absence of other young men in the family, she would have chosen a more suitable occupation for a woman. Nevertheless, the example of this woman, who built an entire empire in extremely difficult circumstances, should inspire everyone.
The most important news and the best texts are in our Telegram channel. Subscribe!
Cover photo: Wikimedia Commons
"Too much of something is bad, but too much champagne is always good"
Francis Scott Fitzgerald
"Champagne is the only wine after which a woman remains beautiful"
Madame de Pompadour
Louis Roederer Cristal
Not a well-known type of champagne in our country - but truly unique. This champagne is handmade, from the grape harvest to the labeling. "Armand de Brignac" is especially loved by representatives of musical bohemians - stars who are used to not denying themselves anything. As Kim Kardashian once said: “If I have to spend money on something, I will first think about this champagne!” (it was “Armand de Brignac” that guests were treated to at her wedding in 2011). Victoria Beckham explains her choice of favorite champagne simply: "It's as chic as me!" ("Posh Spice", that is, "chic", was her nickname in the Spice Girls group). And Oprah Winfrey was hooked on Armand de Brignac by Jay Z - he once sent her a bottle as a gift, and since then the famous presenter knows what to give to all her friends for Christmas. Well, Leonardo DiCaprio: he is such a big fan of Armand de Brignac that on his last birthday he spent 3 million (!) Only on bottles of this wine. This, by the way, does not mean that champagne flowed like water at the party: the price of a bottle of Armand de Brignac of a good year reaches 250 thousand dollars.
Dom Perignon
Fans: James Bond,
"He who drinks '52 Dom Perignon cannot be a bad person" - these words belong to agent 007. Let's not be too harsh: champagne can go to the head of even the best of spies. “Dom Perignon”, in fact, is loved by everyone: gangsters, members of the royal family, and mere mortals. This is a classic of the genre, like a Verdi opera and a little black dress from Chanel. To be fair, James Bond himself has repeatedly cheated on Dom Perignon with competing brands such as Bollinger and Taittinger (depending on who was more generous with product placement). Only in the last films with the participation of Daniel Craig did Bond switch to beer, the drink of the proletariat - what a shame!
Moët & Chandon
Fans:, Claire Danes, Sharon Osbourne, Prince Harry
Someone will think: well, of course, she was his “face”! But, according to the producers of "Moet ...", they chose Scarlett because she repeatedly confessed her love for this variety, and they simply helped her "share this passion." Well, Prince Harry somehow took advantage of his position to treat himself and his company for free with a dozen bottles of Moet & Chandon: an official letter of protest soon arrived from Buckingham Palace: no, their "boy" simply could not drink so much dry sparkling!
The most important and most famous quality of champagne wines is the bubbles, which, bursting, form a small fragrant firework above the glass. Researchers from the birthplace of champagne - from the University of Reims (Champagne, France) - conducted the most accurate mass spectrometric analysis of the substances included in the aerosol that appears above the surface of the sparkling drink. According to the results of the analysis, this aerosol is many times enriched (compared to the liquid phase) with hundreds of aromatic substances that determine the smell of wine, largely thanks to which champagne has won its fame as a noble drink.
Surprisingly tasty, sparkling and spicy!
I'm all in something Norwegian! I'm all in something Spanish!
I get inspired impulsively! And take up the pen!
The sound of airplanes! Run cars!
Express whistle! The winglet of the buoys!
Someone's been kissed here! Someone was killed there!
Pineapple in champagne is the pulse of the evenings!
In a group of nervous girls, in an acute society of ladies
I will turn the tragedy of life into a dream farce...
Pineapples in champagne! Pineapples in champagne!
From Moscow to Nagasaki! From New York to Mars!
I must say that the conceptual predecessor of this study was also work on splashes, but not champagne, but sea water. It has long been established that the sea air (compared to the water column) is many times enriched with organic molecules of marine origin. The mechanism of this phenomenon is quite simple: all these compounds are surfactants, - that is, substances with surface activity , - and, due to their amphiphilic chemical nature, are adsorbed on the surface of bubbles arising in sea waves. Floating to the surface, the bubbles burst, and, breaking up into myriads of microscopic drops, form spray can enriched with these organic molecules.
With champagne, the situation is approximately the same. If this drink is desacralized and guided only by the principles of scientific knowledge, this wine (and other effervescent wines) can be represented as a multicomponent water-alcohol solution supersaturated with carbon dioxide (CO 2 ), which is formed in parallel with alcohol during the fermentation process. However, the most important thing here is not this, but the content of hundreds of surface-active compounds "inherited" from grape raw materials or microorganisms that carry out the entire process. (By the way, a typical bottle of champagne (0.75 liters) contains about 5 liters of CO 2 , which, given the typical bubble size (0.5 mm), adds up to a surface area of about 80 m 2 .)
Every second, the playing wine sprays whole clouds of microscopic droplets that appear after the next emerging gas bubble in the glass bursts. In order not to rely solely on our own organs of vision, this fascinating process has been studied in sufficient detail using high-speed macro photography and laser tomography (Fig. 1).
Figure 1. The process of aerosol formation over the surface of a champagne glass. A - A series of photographs with a time interval of ≈1 ms, illustrating the final stage of the existence of a single bubble (mark: 1 mm). B - Merging with a friend and bursting, champagne bubbles actually raise into the air (in the form of an aerosol) the top layer of liquid. Myriads of microscopic droplets, sprayed in multitudes every second, scatter several centimeters above the surface. V - Aerosol from champagne over the surface of the glass, as it looks with the help of laser tomography techniques.
To study the composition of the aerosol, a glass slide was placed on a glass of champagne for 10 minutes, samples of the settled liquid from which were subjected to mass spectrometric analysis. Comparison of the mass spectra of the aerosol and the liquid phase in the mass-to-charge ratio (m/z) range of 150–1000 revealed thousands of “common” compounds, as well as more than a hundred molecules, the content of which in the aerosol turned out to be several orders of magnitude higher than in the liquid.
To identify these molecules, the scientists searched metabolic databases with an interface for mass spectrometric data, identifying grape metabolites as potential candidates ( Vitis vinifera) and yeast ( Saccharomyces cerevisiae), which are most directly related to the biochemistry of wine. Among the 163 aerosol enriching compounds, 32 are believed to be from grapes and 13 from yeast.
Among the "recognized" molecules in champagne splashes are saturated and unsaturated fatty acids with chain lengths C 13 -C 24, a group of norisoprenoids (terpenes), which determine both the general "outlines" of the smell of wine, and aromas specific to Shiraz, Chardonnay, melon, nutmeg, riesling, and other substances, as a rule, with a characteristic odor.
Gerard Liger-Bélard, who led the team of French and German scientists who did this work, commented on his heightened interest in what was happening in a glass of champagne: "Thanks to these amazing processes, one glass contains both food for the mind and pleasure for the senses".
Literature
- G. Liger-Belair, C. Cilindre, R. D. Gougeon, M. Lucio, I. Gebefugi, et. al. (2009). Unraveling different chemical fingerprints between a champagne wine and its aerosols . Proceedings of the National Academy of Sciences. 106 , 16545-16549;
- Colin D O "Dowd, Gerrit de Leeuw. (2007). Marine aerosol production: a review of the current knowledge . Philosophical Transactions of the Royal Society A: Mathematical, Physical and Engineering Sciences. 365 , 1753-1774;
- Liger-Belair G., Lemaresquier H., Robillard B., Duteurtre B., Jeandet P. (2001). The secrets of fizz in champagne wines: A phenomenological study. Am. J Enol. Vitic. 52 , 88–92;
- Gerard Liger-Belair, Guillaume Polidori, Philippe Jeandet. (2009). ChemInform Abstract: Recent Advances in the Science of Champagne Bubbles. ChemInform. 40 .
We, women, are ready for a lot to find the perfect figure. Sometimes, all kinds of efforts do not help to make the waist narrow and the hips slender. Then we dream of a wonderful way that will help to remove the excess exactly where it is needed, and make the figure chiseled and light. There is such a wonderful way - this is cavitation. And here, as in the story of the birth of the beautiful Aphrodite from the foam, it will not do without magic bubbles.
Cavitation is a method of dealing with local fat deposits. If you want to solve the problem of excess weight, when there is a systemic deposition of fat, then first of all, complex measures are needed that will help speed up the metabolism and start the process of burning fat. You will have to change your diet, increase physical activity, and maybe even change your lifestyle. And only after that, when the total weight has decreased, but dissatisfaction with problem areas remains, the cavitation method comes to the rescue.
“Ultrasonic and EWATage (evacuation) cavitation are distinguished by the nature of the impact,” says Svetlana Nekrasova, a physiotherapist, chief physician of the Aesthetic Medical Center. The essence of ultrasonic exposure is that a wave of a strictly defined length (from 34 to 73 MHz) penetrates tissues and causes swaying of fat cells - a kind of micromassage. As a result of such rocking, microbubbles are formed inside the cell. Their appearance in adipose tissue is the effect of cavitation.”
After the cavitation procedure, you need to eat right and follow the doctor's prescriptions, then the effect of volume loss can reach four centimeters per week.
Then the following happens: the bubbles overflow the cell from the inside, expand it, forming cracks in the membrane. Through them, the contents leave the cell, and then the body. After the procedure, dietary nutrition, lymphatic drainage and a special drinking regimen are recommended.
Evatage-cavitation is even more intense than ultrasonic evacuation. It is based on extracorporeal shock wave therapy, that is, a mechanical effect on tissues. To describe this method, a comparison can be made with how a thrown stone causes circles in the water. Similarly, a series of wave impacts causes destabilization of the adipose tissue. As a result, bubbles also appear, and the content of the cell changes in consistency. In the normal state, the fat cell is quite dense, and cavitation makes it labile and ready for removal. It is wrong to expect a noticeable result immediately after the procedure: excess fat should be removed gradually, then it is a more natural, physiological process.
Both procedures are tolerated comfortably: with ultrasonic cavitation, heat is felt, with evacuation-cavitation - a slight tapping. Which method is more suitable in each case, the doctor decides - the choice depends on the quantity and quality of adipose tissue and other individual characteristics.
“I would like to note,” continues Svetlana Vladimirovna, “that evacuation-cavitation is a development of our center. The manufacturer of shock wave equipment did not provide for its use in order to reduce body fat. But the selection of the necessary nozzle, the calculation of the power and frequency of exposure allowed us to achieve excellent results in this area of application. The results were recognized by the equipment manufacturer, and now we are ready to share our experience at the international level, since there is a great interest in the development of this area in modern cosmetology.”
Photo: Fusion of a vesicle with a cell membrane: “sausages” show SNARE receptor proteins (right) and viral proteins that mimic their work (left).
This year's Nobel Prize in Physiology or Medicine was awarded to three American scientists for "research on the mechanisms that regulate vesicular transport." Randy Shekman, James Rothman and Thomas Zudof in their works explained how various substances move inside cells in membrane vesicles: what genes are needed for this to work, how vesicle fusion occurs at the molecular level, and how this process is regulated in neurons, where it is especially important that the merger happened only at the right time and in the right place.
A eukaryotic, that is, a cell containing a nucleus, from the point of view of biochemistry, is very large. Although it can usually only be seen through a microscope (eggs and orange fibers do not count), even the smallest eukaryotic cell is hundreds and thousands of times larger than a bacterial cell. As complex as a bacterium is, it is ultimately not far removed from a test tube with a (very complex) solution, but eukaryotic cells are very different from nuclear-free microbes in this respect. They are always divided into many departments that perform different functions and often contain completely different, incompatible substances.
This means that eukaryotes, unlike bacteria, faced the problem of intracellular logistics at some point in their evolution. Before nuclear organisms arose, there was no such problem: what was synthesized in one part of the bacterial cell immediately diffused into another part of it. If any substance needed to be thrown into the environment, it was usually synthesized on the membrane, while being pulled out like a thread through the eye of a needle.
However, for a large and complex eukaryotic cell, even if it is a completely independent organism, it is impossible to do without an intracellular transport system. And even more so, such a system is necessary for multicellular organisms, some of whose cells specialize in the production of various substances: hormones, digestive enzymes, or neurotransmitters. That is why eukaryotes, along with the nucleus and mitochondria, have another fundamental innovation - a developed system for the transport of substances in membrane vesicles.
Randy Shekman: From Bubbles to Genes
It should immediately be noted that the current Nobel Prize was awarded not for the discovery of vesicular transport as such, but for elucidating the mechanism of its work. The fact that certain substances can be transported inside cells in container vesicles became clear almost at the same time that the electron microscope became widespread - such vesicles were clearly visible in the photographs. One of the "logistic nodes" where they are formed, the Golgi apparatus, was discovered by the Italian scientist Camillo Golgi at the end of the 19th century, even before the invention of the electron microscope. The second main "cellular hub", the endoplasmic reticulum (EPR), was discovered somewhat later by Albert Claude, for which the scientist, along with two colleagues, received the Nobel Prize in Physiology or Medicine in 1974. And, finally, the fact that it is the membrane vesicles with neurotransmitters that transmit signals from one neuron to another in synapses was established by Katz, von Euler and Axelrod, thanks to which they also became Nobel laureates in 1970.
However, what exactly controls the membrane vesicles, due to which they are transported to the right parts of the cell, how they merge with the cell membrane, remained unclear until the end of the seventies of the last century, when Randy Shekman, a researcher at the University of Berkeley, addressed this issue.
Shekman's supervisor at the university is Artur Kornberg, a Nobel laureate and renowned biochemist (and also the father of Nobel laureate Roger Kornberg, who now chairs the Skolkovo Science Council with Zhores Alferov).
Despite the biochemical school, in order to deal with vesicular transport, Shekman turned not to the biochemical, but to the genetic method of research. He decided to use the simplest eukaryotic model organism, and set about obtaining yeast mutants that exhibit certain defects in vesicular transport.
In a series of works carried out jointly with Peter Novik (it is he who is listed as the first author of Shekman's key articles), the scientist discovered 23 genes in yeast, the work of which is necessary for the normal secretion of glycoproteins. When the mutant yeast was transferred to a thermostat with a high temperature (there the mutations began to show their effect), the cells stopped dividing. Under an electron microscope, thousands of small bubbles could be seen along the edges of such cells, which could not merge with the membrane and throw their contents out. The genes corrupted in these mutants are named sec1,sec2,sec3 etc. They became a kind of library that subsequent scientists were guided by when they began to look for related genes in higher eukaryotes. However, how the proteins encoded by these genes work at the molecular level, it was no longer Shackman who managed to find out, but his independently working colleague, James Rothman.
James Rothman: Protein Lightning
James Rothman, who is only two years younger than Shekman and was working on intracellular transport at Stanford around the same time, had a fundamentally different approach to research. First, he did not work on yeast, but on mammalian cell cultures. More precisely, not even on the cells themselves, but on their extracts. Secondly, he was not engaged in the search for mutants, but in classical biochemical work - the isolation of proteins. In a sense, one can say that Rothman began to "dig the tunnel from the other end" and, fortunately, in 1992, these two lines of research converged in one joint work.
box#1427496
Rothman's main model was the vesicular stomatitis virus (VSV), one of the proteins of which is glycosylated upon maturation, that is, it is modified by various sugars. As this protein, after being synthesized on the ER membrane, moves along the transport "conveyor" of the cell, it first receives and then loses some sugars. These sugars turned out to be very convenient markers for Rothman, thanks to which it was possible to track at what stage the transport stopped when certain cell extracts were added.
Working with this biochemical system, Rothman isolated first one (NSF), and then many proteins, the work of which was necessary for the fusion and division of membrane vesicles. And at this point, the work of Shekman and Rothman, a genetic and biochemical approach, converged: it turned out that one of the proteins isolated from cell extracts (SNAP) is a close relative of the one whose sequence is encoded by the gene sec17 from yeast. The discovery was published in the first joint work of the current Nobel laureates, who up to this point worked completely independently of each other. Among other things, it followed from this coincidence that the vesicular transport system in yeast and mammals works through the same general mechanisms.
Further biochemical experiments by Rotman made it possible to establish the composition of a whole complex of proteins that are involved in the fusion of molecular bubbles. To search for these molecules, the scientist no longer used extracts of cell cultures (there is usually quite a bit of material in them), but preparations of bovine brains, because it is in the nervous tissue that there are a lot of synapses where vesicles with neurotransmitters must merge at the command of electrical excitation.
Rothman's work led to the so-called SNARE hypothesis, a model that explains why vesicles fuse with cell membranes exactly where they are needed. According to this model, fusion is regulated by two groups of receptors: t-(target)-SNARE (syntaxins) and v-(vesicle)-SNARE (synaptobrevins), that is, molecules located on the membrane and on vesicles, respectively. Certain v-SNARE receptors are only able to interact with t-SNARE receptors of exactly the right type (of which there are at least 35 known varieties), so the fusion is specific, although the mechanism is broadly the same.
The key point of fusion is the interweaving of proteins located on different membranes into peculiar braids of four alpha-helices (in the English-language literature they are commonly called "zippers"). This entanglement provides the energy needed to fuse the lipid layers, which normally repel each other quite strongly due to the negative charge of the phosphates.
Thomas Südof: Calcium regulation
After the molecular mechanism of membrane vesicle fusion was elucidated, the question of the temporal regulation of this process remained. Indeed, in nerve cells, vesicles with a neurotransmitter should be ejected into the synaptic cleft if and only if the cell is excited. The electrical depolarization of the neuron always accompanies the entry of calcium ions into the cells, and it was they that turned out to be the key to the whole process.
It was Thomas Südof, a biochemist from Göttingen, who completed his main work already in the USA, at the University of Texas, who managed to establish the details of calcium regulation. He found that, in addition to SNARE receptors, several other proteins play an important role in the process of fusion of membrane vesicles in synapses, the key of which turned out to be complexin and synaptotagmin.
Working on the so-called knockout mice - animals in which one of the genes is artificially turned off, Zudof showed that the removal of complexin leads to a strong decrease in the activity of all synapses without exception. The direct binding of calcium ions is carried out by another protein, synaptotagmin. In addition, Zudof and colleagues found a third protein that corresponded to the same mutant sec-1, which was first caught by Shekman in his research in the late 70s.
Image: Danko Dimchev Georgiev, M.D.
Interestingly, in the course of these experiments, Zudof even managed to obtain a line of knockout mice, in which, due to the absence of one of the proteins, not a single (!) synapse worked in the entire nervous system. The most surprising thing was that such rodents formed an almost normal brain, the neurons of which still die, but very late - only after its full maturation. Thus, along with the clarification of the details of the regulation of vesicular transport, it was possible to establish that the work of synapses is necessary for the brain in order to maintain its existence, but is not required until it has matured yet.
About the fashion for science
Last year's Nobel Prize in Medicine was awarded to John Gardon and Shinya Yamanaka for their discovery of a reprogramming mechanism that makes it possible to obtain stem cells from almost any mature cell. The works of these two scientists were greatly spaced in time - Gardon conducted key experiments in the 70s, and Yamanaka received the first reprogrammed stem cells in 2004. To say that this latest discovery has been highly anticipated is an understatement: it finally made it possible to work with stem cells without the use of embryos and, more importantly, taught biologists to obtain stem cells that are genetically identical to the material donors. Today, such cells are already being used with might and main to obtain artificial organs. Of these, as scientists have recently shown, even brain-like organelles are formed, and produced in situ, such cells have full totipotency - they are even able to form embryos inside the body.
Vesicular transport, compared to cellular reprogramming, seems to be a much less “fashionable” topic. Perhaps such an alternation of fashionable and not too fashionable topics is a conscious policy of the Nobel Committee, or maybe just the result of chance. In any case, the Stockholm experts are still unpredictable: none of the topics that were promised this year's medicine award won. But among them there was such an important topic as epigenetic methylation - it was on it that many people in the molecular biological community "bet".
The Nobel Committee, as we see, does not always follow fashion. And this is good: in the long run, the value of a discovery is determined not by its immediate applicability, but by its fundamental nature, that is, by how deep processes it can explain.
If someone really wants to give the current award a fashionable flair, then this is as easy as shelling pears. Remember such a cosmetic procedure as "Botox", an injection of botulinum toxin? So, botulinum toxin cuts exactly the same proteins discovered by Rothman (namely SNAP-25) in the SNARE receptor complex at the vesicle fusion site, which leads to the shutdown of this synapse.
