4NIV

Crystal structure of trypsiligase (K60E/N143H/Y151H/D189K trypsin) trigonal form

4NIV の概要

• エントリーDOI: 10.2210/pdb4niv/pdb
• 関連するPDBエントリー: 4NIW 4NIX 4NIY
• 分子名称: Cationic trypsin, CALCIUM ION, GLYCEROL, ... (4 entities in total)
• 機能のキーワード: trypsin, serine proteinase, enzyme design, activation domain, zymogen, peptide ligation, reverse proteolysis, hydrolase
• 由来する生物種: Bos taurus (bovine,cow,domestic cattle,domestic cow)
• 細胞内の位置: Secreted, extracellular space: P00760
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 23469.50

構造登録者

Schoepfel, M.,Parthier, C.,Stubbs, M.T. (登録日: 2013-11-08, 公開日: 2014-02-19, 最終更新日: 2024-11-27)

主引用文献

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

実験手法

X-RAY DIFFRACTION (1 Å)