PRIMARY STRUCTURE REQUIREMENTS FOR THE MATURATION INVIVO OF PENICILLIN ACYLASE FROM ESCHERICHIA-COLI ATCC-11105

被引:86
作者
SIZMANN, D [1 ]
KEILMANN, C [1 ]
BOCK, A [1 ]
机构
[1] UNIV MUNICH, LEHRSTUHL MIKROBIOL, MARIA WARD STR 1A, W-8000 MUNICH 19, GERMANY
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1990年 / 192卷 / 01期
关键词
D O I
10.1111/j.1432-1033.1990.tb19207.x
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
The two constituent subunits of the enzyme penicillin acylase from Escherichia coli strain ATCC 11105 are derived from a single precursor polypeptide by post‐translational processing. Mutant penicillin acylase precursors were constructed carrying insertions and deletions in various domains and they were analysed for their processing behaviour. It was found that an endopeptide region of appropriate size and an intact C‐terminus were absolutely necessary for the maturation process. Internal deletions within the β‐subunit domain also prevented post‐translational cleavage. Processing competence, therefore, was not merely determined by the amino acid sequence in the vicinity of the processing sites but relied on a correct overall conformation of the protein. The processing pathway in vivo proceeds via an intermediate comprising the α subunits plus endopeptide and is thus identical to the pathway which has been determined previously by in vitro analysis. The post‐translational modification of the precursor is probably not carried out by a specific processing enzyme(s) as the heterologous expression of the penicillin acylase (pac) structural gene yielded processed and active enzyme in different enterobacteria and in a Pseudomonas species. Copyright © 1990, Wiley Blackwell. All rights reserved
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页码:143 / 151
页数:9
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共 37 条
  • [31] SIZMANN D, 1985, THESIS MUNICH U
  • [32] SIZMANN D, 1989, THESIS MUNICH U
  • [33] TANIUCHI H, 1970, J BIOL CHEM, V245, P5459
  • [34] TANIUCHI H, 1969, J BIOL CHEM, V244, P3864
  • [35] AN UNUSUAL GENE-CLUSTER FOR THE CYTOCHROME-BC1 COMPLEX IN BRADYRHIZOBIUM JAPONICUM AND ITS REQUIREMENT FOR EFFECTIVE ROOT NODULE SYMBIOSIS
    THONYMEYER, L
    STAX, D
    HENNECKE, H
    [J]. CELL, 1989, 57 (04) : 683 - 697
  • [36] A SINGLE GENE DIRECTS SYNTHESIS OF A PRECURSOR PROTEIN WITH BETA-AMYLASE AND ALPHA-AMYLASE ACTIVITIES IN BACILLUS-POLYMYXA
    UOZUMI, N
    SAKURAI, K
    SASAKI, T
    TAKEKAWA, S
    YAMAGATA, H
    TSUKAGOSHI, N
    UDAKA, S
    [J]. JOURNAL OF BACTERIOLOGY, 1989, 171 (01) : 375 - 382
  • [37] PRO-SEQUENCE OF SUBTILISIN CAN GUIDE THE REFOLDING OF DENATURED SUBTILISIN IN AN INTERMOLECULAR PROCESS
    ZHU, XL
    OHTA, Y
    JORDAN, F
    INOUYE, M
    [J]. NATURE, 1989, 339 (6224) : 483 - 484