Trypsin complexed with α 1‐proteinase inhibitor has an increased structural flexibility

FEBS Letters

Patthy, András¹  Gráf, László²  Kaslik, Gyula²  Bálint, Miklós² 

[1]Agricultural Biotechnology Center, P.O.B. 170, H-2100 Gödöllõ, Hungary

[2]Department of Biochemistry, Eötvös University, Puskin u. 3, H-1088 Budapest, Hungary

关键词: α 1-Antitrypsin;    Mutant rat trypsin;    Acyl enzyme intermediate;    Limited proteolysis;    Induced fit;    α 1-PI;    α 1-proteinase inhibitor;    TPCK;    N-tosyl-l-phenylalanine-chloromethyl ketone;    TFA;    trifluoroacetic acid;    PMSF;    phenylmethylsulfonyl fluoride;    SDS;    sodium dodecyl sulfate;    PAGE;    polyacrylamide gel electrophoresis;    FPLC;    fast protein liquid chromatography;    HPLC;    high pressure liquid chromatography;   

DOI  :  10.1016/0014-5793(95)00816-R

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Mutant rat trypsin Asp¹⁸⁹Ser was prepared and complexed with highly purified human α ₁-proteinase inhibitor. The complex formed was purified to homogeneity and studied by N-terminal amino acid sequence analysis and limited proteolysis with bovine trypsin. As compared to uncomplexed mutant trypsin, the mutant enzyme complexed with α ₁-proteinase inhibitor showed a highly increased susceptibility to enzymatic digestion. The peptide bond selectively attacked by bovine trypsin was identified as the Arg¹¹⁷-Val¹¹⁸ one of trypsin. The structural and mechanistic relevance of this observation to serine proteinase-substrate and serine proteinase-serpin reactions are discussed.

【 授权许可】

Unknown