Chemical modification of porcine kidney aminopeptidase P indicates the involvement of two critical histidine residues

FEBS Letters

Lim, Jaeseung¹  Turner, Anthony J.¹ 

[1] Department of Biochemistry and Molecular Biology, University of Leeds, Leeds LS2 9JT, UK

关键词: Aminopeptidase P;    Diethylpyrocarbonate;    Metallopeptidase;    Proline-peptidase;    Histidine modification;    Porcine kidney;    AP;    aminopeptidase;    DEP;    diethylpyrocarbonate;    GPI;    glycosylphosphatidylinositol;    Hyp;    hydroxyproline;    PLC;    phospholipase C;   

DOI  :  10.1016/0014-5793(96)00124-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Aminopeptidase P (AP-P), purified to homogeneity from porcine kidney membranes, was completely inactivated by treatment with 0.2 mM diethylpyrocarbonate (DEP) at pH 7.0. Treatment of the modified enzyme with 20 mM hydroxylamine resulted in recovery of AP-P activity. The differential absorption of native and modified AP-P at 240 nm showed that DEP modified two histidyl residues per mol of AP-P. The substrates, bradykinin(1–5) and Gly-Pro-Hyp, and also the inhibitor, apstatin, could protect against DEP inactivation. These results suggest that histidine residues are critical for AP-P activity.

【 授权许可】

Unknown