A PUTATIVE METAL-BINDING SITE IN THE BETA-SUBUNIT OF RAT MITOCHONDRIAL PROCESSING PEPTIDASE IS ESSENTIAL FOR ITS CATALYTIC ACTIVITY

被引：53

作者：

KITADA, S ^{[1
]}

SHIMOKATA, K ^{[1
]}

NIIDOME, T ^{[1
]}

OGISHIMA, T ^{[1
]}

ITO, A ^{[1
]}

机构：

[1] KYUSHU UNIV,FAC SCI,DEPT CHEM,HIGASHI KU,FUKUOKA 812,JAPAN

来源：

JOURNAL OF BIOCHEMISTRY | 1995年 / 117卷 / 06期

关键词：

METALLOENDOPEPTIDASE; MITOCHONDRIAL PROCESSING PEPTIDASE; PROTEIN EXPRESSION; SITE-DIRECTED MUTAGENESIS;

D O I：

10.1093/oxfordjournals.jbchem.a124836

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Mitochondrial processing peptidase (MPP) consists of alpha- and beta-subunits (alpha-MPP and beta-MPP). beta-MPP has a putative metal-binding sequence (HXXEH). To determine whether the sequence of beta-MPP is essential for the enzymatic activity, we individually mutated the histidines and glutamic acid to arginines and glutamine, respectively. The wild-type and mutated beta-MPPs were co-expressed with alpha-MPP in Escherichia coli. All three mutants had completely lost the activity, whereas the lost activity was recovered on the addition of wild-type beta-MPP. The activity of the wild-type enzyme was reduced by the mutant beta-MPPs. We conclude from these observations that the HXXEH region is involved in the formation of the active site and that beta-MPP is the catalytic subunit of MPP.

引用

页码：1148 / 1150

页数：3

共 20 条

[1]

ARRETZ M, 1994, J BIOL CHEM, V269, P4959

[2] BIOGENESIS OF MITOCHONDRIA [J].

ATTARDI, G ;

SCHATZ, G .

ANNUAL REVIEW OF CELL BIOLOGY, 1988, 4 :289-333

[3] AN UNUSUAL ACTIVE-SITE IDENTIFIED IN A FAMILY OF ZINC METALLOENDOPEPTIDASES [J].

BECKER, AB ;

ROTH, RA .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (09) :3835-3839

[4] THE GENERAL MITOCHONDRIAL PROCESSING PEPTIDASE FROM POTATO IS AN INTEGRAL-PART OF CYTOCHROME-C REDUCTASE OF THE RESPIRATORY-CHAIN [J].

BRAUN, HP ;

EMMERMANN, M ;

KRUFT, V ;

SCHMITZ, UK .

EMBO JOURNAL, 1992, 11 (09) :3219-3227

[5]

EMMERMANN M, 1993, J BIOL CHEM, V268, P18936

[6] FUNCTIONAL RECONSTITUTION IN ESCHERICHIA-COLI OF THE YEAST MITOCHONDRIAL MATRIX PEPTIDASE FROM ITS 2 INACTIVE SUBUNITS [J].

GELI, V .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (13) :6247-6251

[7] MITOCHONDRIAL PROTEIN IMPORT [J].

HARTL, FU ;

PFANNER, N ;

NICHOLSON, DW ;

NEUPERT, W .

BIOCHIMICA ET BIOPHYSICA ACTA, 1989, 988 (01) :1-45

[8] MITOCHONDRIAL PROTEIN IMPORT - IDENTIFICATION OF PROCESSING PEPTIDASE AND OF PEP, A PROCESSING ENHANCING PROTEIN [J].

HAWLITSCHEK, G ;

SCHNEIDER, H ;

SCHMIDT, B ;

TROPSCHUG, M ;

HARTL, FU ;

NEUPERT, W .

CELL, 1988, 53 (05) :795-806

[9] IMPORT OF PROTEINS INTO YEAST MITOCHONDRIA - THE NUCLEAR MAS2 GENE ENCODES A COMPONENT OF THE PROCESSING PROTEASE THAT IS HOMOLOGOUS TO THE MAS1-ENCODED SUBUNIT [J].

JENSEN, RE ;

YAFFE, MP .

EMBO JOURNAL, 1988, 7 (12) :3863-3871

[10] 2 MITOCHONDRIAL MATRIX PROTEASES ACT SEQUENTIALLY IN THE PROCESSING OF MAMMALIAN MATRIX ENZYMES [J].

KALOUSEK, F ;

HENDRICK, JP ;

ROSENBERG, LE .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (20) :7536-7540

← 1 2 →