Mapping the stereospecificity of peptidyl prolyl cis/trans isomerases

FEBS Letters

Reimer, Ulf¹  Schutkowski, Mike¹  Schiene, Cordelia¹  Fischer, Gunter¹ 

[1] Max-Planck Society, Research Unit ‘Enzymology of Protein Folding', Kurt-Mothes-Str. 3, D-06120 Halle/Saale, Germany

关键词: Peptidyl prolyl cis/trans isomerase;    Stereospecificity;    Proline;    Cis/trans isomerization;    Enzyme mechanism;    Abz;    2-aminobenzoyl residue;    CsA;    cyclosporin A;    Cyp18;    recombinant human cytosolic cyclophilin with molecular mass of 18 kDa;    DMSO;    dimethyl sulfoxide;    FKBP;    FK506 binding protein;    HPLC;    high performance liquid chromatography;    -NH-Np;    4-nitroanilide;    PPIase;    peptidyl prolyl cis/trans isomerase;    TF domain;    Escherichia coli trigger factor PPIase domain (segment Arg145 to Glu251);    TLC;    thin layer chromatography;   

DOI  :  10.1016/S0014-5793(98)00871-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The stereospecificity of peptidyl prolyl cis/trans isomerases (PPIases) was studied using tetrapeptide substrate analogs in which one amino acid residue was replaced by the cognate d-amino acid in various positions of the peptide chain. Reversed stereocenters around proline markedly increased the rate of the spontaneous trans to cis isomerization of the prolyl bond whereas cis to trans isomerizations were less sensitive. PPIases like human cyclophilin18, human FKBP12, Escherichia coli parvulin10 and the PPIase domain of E. coli trigger factor exhibited stereoselectivity demanding at the P₁ to P₂′ position of the substrate chain. The discriminating factor for stereoselectivity was the lack of formation of the Michaelis complexes of the diastereomeric substrates. However, d-alanine at the P₁ position preserved considerable affinity to the active site, and largely prevented activation of the catalytic machinery for all PPIases investigated.

【 授权许可】

Unknown