Native chemical ligation (NCL) is a useful method for synthetic and semi-synthetic proteins synthesis. It is excited that a synthetic component in NCL can be site-specifically glycosylated, retaining native sugar-protein linkages. This technology allows the assembly of glycoproteins that are not readily available from biological sources. With extensive experience in employing NCL for glycoprotein synthesis, scientists of Creative Biolabs are proficient in synthesizing glycoprotein up to 150 amino acids in length.
Glycoproteins are featured by a covalent linkage between carbohydrates through asparagine and serine or threonine. The carbohydrates are usually N-acetyl glucosamine, N-acetyl galactosamine, mannose, xylose, and fucose. Sufficient quantities of homogeneous glycoproteins are often not available from biological sources to facilitate glycoprotein research. Previous studies have shown that the total chemical synthesis of certain glycopeptide components of glycoproteins is possible. Indeed, the difficulty associated with glycoprotein synthesis has precluded much biological research using entirely chemical approaches.
In the process of NCL, a C-terminal peptide thioester reacts with an N-terminal cysteinyl peptide to produce a native peptide bond between the two fragments, which is convenient to use NCL for the synthesis of active proteins from smaller fragments. Such an approach allows protein modifications (including glycosylation, phosphorylation, acetylation, methylation, and sulfation) to be introduced in a controlled fashion into smaller peptide fragments that are amenable to total chemical synthesis. With defined sequence and structure, these fragments can be ligated into full-length proteins. NCL allows controlling the identity and the position of glycosylation, which is only limited by the imagination and capability of the synthetic chemist, making NCL the most popular chemical approach for glycoprotein synthesis.
Fig.1 The mechanism of NCL. (Thapa, 2014)
It is widely accepted that approximately 50 amino acid residues represent the limit for efficient automated synthesis and the difficulties are associated with glycoprotein synthesis. To address this, Creative Biolabs has introduced a group of scientists who have a distinguished understanding of glycoprotein synthesis. NCL has greatly extended the size of proteins that can be synthesized. Based on NCL, we have successfully synthesized glycoproteins up to 150 amino acids in length. We are fully competent and dedicated to serving as your one-stop-shop for glycoprotein synthesis and characterization.
Equipped with professional scientific staff and world-leading technology platforms in glycoprotein research, Creative Biolabs is pleased to share our cutting-edge technology and extensive expertise in using NCL for glycoprotein synthesis. If you are interested in glycoprotein synthesis, please contact us for more information and a detailed quote.