Russian scientists use coral fluorochromes to identify proteins

This two-color label is called Dendra, which is derived from the Dendronephthya. Under the microscope, we can observe that the two-color label first emits green light, and identifies its attached proteins that are invisible to the naked eye. When the tag receives a visible blue light pulse, the green light turns red. Constantin Lukyanov said that this color change "makes it possible to use a single light to accurately identify the same entity, such as a cell, a cell organ, or a protein, and then track its stroke in real time." He completed the Dendra experiment in his brother's lab at Seiergai Lukjanov. Shirgege is an international research scholar at the Institute of Bioorganic Chemistry, Shayarkin-Voznikov, Moscow State Academy. Constantin said: "This new tool offers us the possibility to study protein and cell organ motility in living cells, cell movement during inflammation, inflammation, and other morbid or normal processes."

Constantin explained that Dendra, unlike other green-red fluorescent markers, is activated by visible blue light, which is less harmful to living cells and does not require special laser instruments. Sylgai believes that the fact that the light emitted by the laser scanning confocal microscope, which is commonly used when observing living cells, can activate the label, so that this new tool can help more scientists. The research results were published in the "Natural" biotechnical edition published in April 2006 and the online edition magazine on March 19.

More importantly, Dendra is already small enough to identify proteins without impeding protein folding and function. Researchers have also demonstrated that it can change shades as body temperature changes in birds and mammals, and that birds and mammals are all research models that researchers typically use. In addition, scientists have also found that it can remain red during long-term protein tracking, which further demonstrates its great role.

Douglas-Plough isolated the green fluorescent protein (GFP) gene from a luminous jellyfish in 1992. Then Martin-Chalfil first used GFP to identify objects in 1994. Since then, researcher Roger Frank has developed a series of high-performance GFP mutants. Mather-Chemo, author of the book “Glowing Genes: A Revolution in Biotechnology” (2005) by Computational Chemist at the Connecticut Institute of Technology, said that Dendra is actually an emerging photoelectrically activated fluorescent protein (PAFPs). A new member of the extended family, photoelectrically activating fluorescent proteins will become an innovative image display tool.

Dominic believes that Lukyanov’s first discovery of a protein that resembles GFP in corals has resulted in a major breakthrough in the application of fluorescent proteins. Before that, no one thought of looking for coral-like proteins similar to GFP, because corals do not glow in the dark like fireflies and jellyfish. The coral's own green and red fluorescent proteins emit light only when activated by higher intensity light. New discoveries will help scientists find many new GFP-like proteins on more non-bioluminescent or even non-fluorescent marine organisms.

At the same time, Shirge Gay and other researchers also discovered that certain wavelengths of light can cause some strong visual changes in proteins, they can never emit light, or change color. Later, they found that these GFP-like proteins that were isolated from sea corals had the ability to change color, but for many research applications, the protein was too large. Thus, the Lukjanov brothers and their colleagues began to create a smaller, more functional fluorescent protein. They systematically mutated the DNA sequence encoding the fluorescent protein, and then expressed the mutated protein in bacteria until a protein that could irreversibly change from green to red was found.

Like GFP, Dendra's gene sequence and an attached short amino acid tail can be inserted into the gene encoding the protein the scientist wants to study. In living cells, this fluorescent tail is produced with the protein. For researchers who want to study the function and whereabouts of major proteins, the tail is like a lighthouse. To test how well the new tag was doing in identifying and tracking the target protein, the team fused Dendra to the protein to form a cell scaffold that includes actin and tubulin filaments, and discovered the intracellular The expected pattern of protein distribution.

Dendra belongs to a small family of green-to-red electro-optically activated fluorescent proteins. The first member of the family, the protein Kaede (meaning maple in Japanese), was discovered four years ago by the Japanese scientist, Templeton. Constantine said: "All known proteins similar to Kaede are sensitive to UV radiation, in which case they will immediately change from green to red, but they are not sensitive to blue light." So the researchers were It is not expected that Dendra can be activated by low toxicity visible light. “When we observed that Dendra had a significant image shift under intense blue light, we couldn't believe it was true.” Constantin said, “We still can't explain why Dendra is different from other similar Kaede's fluorescent protein."

Sergiev Lukejanov pointed out that the UV rays that activate many PAFPs are toxic to cells and can even drastically alter their biochemical properties. In addition, instruments that emit ultraviolet light are very expensive and scarce. Shirgege said, "We expect Dendra to expand the application of photo-activated fluorescent proteins (PAFPs)."

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