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How to use NCL technology to synthesize proteins with more than 300 amino acids? What is the key behind this?
In today's biotechnology field, synthesizing more than 300 amino acids of protein has always been the goal of scientists.This not only involves complex techniques in genetic engineering, but also requires new chemical methods to support it.In this regard,
The emergence of Native Chemical Ligation (NCL) provides an effective solution for the synthesis of complex proteins.The core of NCL technology is that it can bind multiple unprotected peptides in a covalent manner to construct long-chain polypeptides or proteins.
Principles and Processes of NCL Technology
The key to the chemical process of NCL lies in its first step in reaction, namely, the dissociated thiol group of the N-terminal cysteine residue reacts with the C-terminal sulfate.This process is usually carried out in the aqueous phase under physiological conditions (pH ~7.0).When the thiol of the N-terminal cysteine reacts with the C-terminal sulfate of the second unprotected peptide, a reversible intermediate is formed.This intermediate is then rapidly recombined through the internal S,N-acyl transfer mechanism, creating the desired amine bond.
The advantage of this technology lies in its specificity and selectivity, which can effectively avoid the generation of by-products.
Key Components of NCL
The most commonly used catalyst in the NCL process is 4-mercaptophenylacetic acid (MPAA), which is essential for improving product yields.Furthermore, NCL has excellent chemoselectivity, which means that there is little reaction with other functional groups in the reaction, which is ideal for synthesis of complex proteins.
Looking at the development of NCL technology from history
The basis of NCL technology stems from the concept of chemical ligation proposed by Stephen Kent and Martina Schnölzer in 1992, the first practical method for the synthesis of unprotected peptides.Then in 1994, Philip Dawson et al. expanded this concept and proposed NCL technology.This breakthrough led to the formation of native amine bonds, which can more effectively synthesize macromolecular proteins.
The excellence of NCL technology lies in its economic atomic utilization rate and environmentally friendly solvent use, demonstrating a strong response to the concept of green chemistry.
The application of NCL in protein synthesis
NCL technology can be widely used in the synthesis of macromolecular proteins, which is crucial for creating enzymes or proteins with special functions.Due to the high efficiency of NCL, this technology makes it feasible to synthesize large amounts of functional proteins that may have great potential in biomedical research and drug development.
Emerging technologies and future prospects
The evolution of NCL technology has also promoted the development of other innovative technologies, such as the SEA (Bis(2-sulfanylethyl)amido) method, which makes the synthesis of complex proteins more flexible and convenient.These variant technologies further expand our ability to synthesize proteins using chemical methods and open up a broader space for exploration.
Conclusion
To sum up, NCL technology not only changes the way proteins are synthesized, it has also had a profound impact on many areas of biological science.With the advancement of science and technology, this method will undoubtedly continue to promote the expansion of scientific boundaries.In the future, how will this technology help the development of a new generation of proteins?
