KININASE ACTIVITY IN HUMAN PLATELETS - CLEAVAGE OF THE ARG1-PRO2 BOND OF BRADYKININ BY AMINOPEPTIDASE-P

被引:35
作者
VANHOOF, G
DEMEESTER, I
GOOSSENS, F
HENDRIKS, D
SCHARPE, S
YARON, A
机构
[1] UNIV INSTELLING ANTWERP,FAC MED,DEPT CLIN BIOCHEM,UNIV PLEIN 1,B-2610 WILRIJK,BELGIUM
[2] WEIZMANN INST SCI,DEPT MEMBRANE RES & BIOPHYS,IL-76100 REHOVOT,ISRAEL
关键词
D O I
10.1016/0006-2952(92)90439-P
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
A proline-specific peptidase aminopeptidase P (APP, EC 3.4.11.9) that cleaves the Arg1-Pro2 bond of bradykinin was isolated from human platelets by liquid chromatography. The enzyme was purified 557 times. The native molecule has a M(r) of 223,000. Human platelet APP exists as a trimer with a subunit M(r) of 71,000. The apparent K(m) of platelet APP is 66-mu-mol/L for bradykinin and 47-mu-mol/L for the internally quenched fluorogenic substrate Lys (2,4-dinitrophenyl)-Pro-Pro-NH-CH2-CH2-NH-2-aminobenzoyl. 2HCl which is used for the routine determination of the enzyme activity. The optimum pH for hydrolysis of the fluorogenic substrate is 8.0, and the optimum temperature is 43-degrees. Platelet APP is inhibited by 1,10-phenanthroline and activated by Mn2+, thus confirming its metalloprotease nature. Cu2+, Zn2+ and Hg2+ are strongly inhibitory. Inhibition by cysteine protease inhibitors suggests the presence of a thiol group essential for enzymatic activity. Serine protease inhibitors do not affect the enzyme activity.
引用
收藏
页码:479 / 487
页数:9
相关论文
共 27 条
[1]  
ACHUTAMURTHY PN, 1985, FED PROC, V44, P876
[2]  
BAKER CRF, 1991, CIRC SHOCK, V33, P47
[3]  
Cornish-Bowden A, 1988, ENZYME KINETICS
[4]   CLEAVAGE OF PROLYL PEPTIDES BY KIDNEY PEPTIDASES - PARTIAL PURIFICATION OF A X-PROLYL-AMINOPEPTIDASE FROM SWINE KIDNEY MICROSOMES [J].
DEHM, P ;
NORDWIG, A .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1970, 17 (02) :364-&
[5]  
Demuth H U, 1989, J Enzyme Inhib, V2, P239, DOI 10.3109/14756368909088477
[6]  
Erdos E., 1979, HDB EXPT PHARM S, VXXV, P427
[7]   SEQUENTIAL HYDROLYSIS OF PROLINE-CONTAINING PEPTIDES WITH IMMOBILIZED AMINOPEPTIDASES [J].
FLEMINGER, G ;
YARON, A .
BIOCHIMICA ET BIOPHYSICA ACTA, 1983, 743 (03) :437-446
[8]   AMINOPEPTIDASE-P FROM RAT-BRAIN - PURIFICATION AND ACTION ON BIOACTIVE PEPTIDES [J].
HARBECK, HT ;
MENTLEIN, R .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1991, 198 (02) :451-458
[9]   AMINOPEPTIDASE-P AND DIPEPTIDYL PEPTIDASE-IV ACTIVITY IN HUMAN-LEUKOCYTES AND IN STIMULATED LYMPHOCYTES [J].
HENDRIKS, D ;
DEMEESTER, I ;
UMIEL, T ;
VANHOOF, G ;
VANSANDE, M ;
SCHARPE, S ;
YARON, A .
CLINICA CHIMICA ACTA, 1991, 196 (2-3) :87-96
[10]   SIMPLIFIED METHOD FOR SILVER STAINING OF PROTEINS IN POLYACRYLAMIDE GELS AND THE MECHANISM OF SILVER STAINING [J].
HEUKESHOVEN, J ;
DERNICK, R .
ELECTROPHORESIS, 1985, 6 (03) :103-112