Investigation of Serine‐Proteinase‐Catalyzed Peptide Splicing in Analogues of Sunflower Trypsin Inhibitor 1 (SFTI‐1) |
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| Authors: | Natalia Karna Dr. Anna Łęgowska Dr. Stanisław Malicki Dr. Dawid Dębowski Przemysław Golik Agata Gitlin Dr. Przemysław Grudnik Dr. Benedykt Wladyka Dr. Krzysztof Brzozowski Dr. Grzegorz Dubin Prof. Krzysztof Rolka |
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| Affiliation: | 1. Department of Bioorganic Chemistry, Faculty of Chemistry, University of Gdansk, Gdansk, Poland;2. Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland;3. Malopolska Centre of Biotechnology, Jagiellonian Univeristy, Krakow, Poland |
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| Abstract: | Serine‐proteinase‐catalyzed peptide splicing was demonstrated in analogues of the trypsin inhibitor SFTI‐1: both single peptides and two‐peptide chains (C‐ and N‐terminal peptide chains linked by a disulfide bridge). In the second series, peptide splicing with catalytic amount of proteinase was observed only when formation of acyl–enzyme intermediate was preceded by hydrolysis of the substrate Lys–Ser peptide bond. Here we demonstrate that with an equimolar amount of the proteinase, splicing occurs in all the two‐peptide‐chain analogues. This conclusion was supported by high resolution crystal structures of selected analogues in complex with trypsin. We showed that the process followed a direct transpeptidation mechanism. Thus, the acyl–enzyme intermediate was formed and was immediately used for a new peptide bond formation; products associated with the hydrolysis of the acyl–enzyme were not observed. The peptide splicing was sequence‐ not structure‐specific. |
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| Keywords: | enzymes inhibitors peptide splicing SFTI-1 |
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