Serine protease inhibitor mediated peptide bond re-synthesis in diverse protein molecules

• Published in: FEBS letters 2011-11, Vol.585 (21), p.3465-3470
• Main Authors: Sharma, Amit, Radha Kishan, K.V.
• Format: Article
• Language: English
• Subjects: Animals; Models, Molecular; Muramidase - chemistry; Muramidase - metabolism; Nicked protein; Peptides - metabolism; Phenylmethylsulfonyl Fluoride - pharmacology; PMSF; Protein Conformation; Proteins - chemistry; Proteins - metabolism; Proteolysis; Proteolysis - drug effects; Religation; Serine Proteinase Inhibitors - pharmacology; Subtilisin Carlsberg; Subtilisins - antagonists & inhibitors; Subtilisins - metabolism
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/j.febslet.2011.10.004

► Protease inhibitor, PMSF mediated peptide re-synthesis. ► Fast religation of peptides in the presence of a small molecule. ► X-ray structure of a nicked protein. Protease inhibitors have been extensively used in research to prevent unwanted degradation of proteins during purification and analysis. Here, we report a remarkable discovery of protease inhibitor mediated reformation of peptide bonds by the serine protease inhibitor, PMSF in a diverse set of proteolyzed molecules. Interestingly, the religation reaction in the presence of PMSF occurs in a very short time period and with very high yields of the religated product. We also investigate the plausible mechanism of such a reaction and demonstrate through biochemical studies and X-ray crystallography that proximity of reacting termini is essential for the feasibility of this reaction.