4NIY
Crystal structure of trypsiligase (K60E/N143H/Y151H/D189K trypsin) complexed to YRH-ecotin (M84Y/M85R/A86H ecotin)
Summary for 4NIY
| Entry DOI | 10.2210/pdb4niy/pdb |
| Related | 4NIV 4NIW 4NIX |
| Descriptor | Cationic trypsin, Ecotin, CALCIUM ION, ... (5 entities in total) |
| Functional Keywords | trypsin inhibitor, serine proteinase, enzyme design, activation domain, peptide ligation, reverse proteolysis, hydrolase-hydrolase inhibitor complex, hydrolase/hydrolase inhibitor |
| Biological source | Bos taurus (bovine,cow,domestic cattle,domestic cow) More |
| Cellular location | Secreted, extracellular space: P00760 Periplasm: P23827 |
| Total number of polymer chains | 8 |
| Total formula weight | 158557.26 |
| Authors | Schoepfel, M.,Parthier, C.,Stubbs, M.T. (deposition date: 2013-11-08, release date: 2014-02-19, Last modification date: 2024-10-30) |
| Primary citation | Liebscher, S.,Schopfel, M.,Aumuller, T.,Sharkhuukhen, A.,Pech, A.,Hoss, E.,Parthier, C.,Jahreis, G.,Stubbs, M.T.,Bordusa, F. N-terminal protein modification by substrate-activated reverse proteolysis. Angew.Chem.Int.Ed.Engl., 53:3024-3028, 2014 Cited by PubMed Abstract: Although site-specific incorporation of artificial functionalities into proteins is an important tool in both basic and applied research, it can be a major challenge to protein chemists. Enzymatic protein modification is an attractive goal due to the inherent regio- and stereoselectivity of enzymes, yet their specificity remains a problem. As a result of the intrinsic reversibility of enzymatic reactions, proteinases can in principle catalyze ligation reactions. While this makes them attractive tools for site-specific protein bioconjugation, competing hydrolysis reactions limits their general use. Here we describe the design and application of a highly specific trypsin variant for the selective modification of N-terminal residues of diverse proteins with various reagents. The modification proceeds quantitatively under native (aqueous) conditions. We show that the variant has a disordered zymogen-like activation domain, effectively suppressing the hydrolysis reaction, which is converted to an active conformation in the presence of appropriate substrates. PubMed: 24520050DOI: 10.1002/anie.201307736 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.84 Å) |
Structure validation
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