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

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 na...

Full description

Saved in:
Bibliographic Details
Published in:FEBS letters 1996-03, Vol.381 (3), p.188-190
Main Authors: Lim, Jaeseung, Turner, Anthony J.
Format: Article
Language:English
Subjects:
Aminopeptidase P
Aminopeptidases - chemistry
Aminopeptidases - isolation & purification
Aminopeptidases - metabolism
Animals
AP, aminopeptidase
Binding Sites
Bradykinin - pharmacology
Cell Membrane - enzymology
DEP, diethylpyrocarbonate
Diethyl Pyrocarbonate - pharmacology
Diethylpyrocarbonate
Enzyme Inhibitors - pharmacology
GPI, glycosylphosphatidylinositol
Histidine
Histidine modification
Hydroxylamine
Hydroxylamines - pharmacology
Hyp, hydroxyproline
Kidney - enzymology
Kinetics
Metallopeptidase
Oligopeptides - pharmacology
Peptides - pharmacology
PLC, phospholipase C
Porcine kidney
Proline-peptidase
Spectrophotometry, Ultraviolet
Swine
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(96)00124-X