4NIX

Crystal structure of trypsiligase (K60E/N143H/Y151H/D189K trypsin) orthorhombic form, zinc-bound

Summary for 4NIX

• Entry DOI: 10.2210/pdb4nix/pdb
• Related: 4NIV 4NIW 4NIY
• Descriptor: Cationic trypsin, CALCIUM ION, GLYCEROL, ... (5 entities in total)
• Functional Keywords: trypsin, serine proteinase, enzyme design, activation domain, zymogen, peptide ligation, reverse proteolysis, hydrolase
• Biological source: Bos taurus (bovine,cow,domestic cattle,domestic cow)
• Cellular location: Secreted, extracellular space: P00760
• Total number of polymer chains: 1
• Total formula weight: 23534.91

Authors

Schoepfel, M.,Parthier, C.,Stubbs, M.T. (deposition date: 2013-11-08, release date: 2014-02-19, Last modification date: 2024-10-16)

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

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: 24520050
DOI: 10.1002/anie.201307736

Experimental method

X-RAY DIFFRACTION (1.3 Å)