A GENETIC-ENGINEERING APPROACH TO STUDY THE MODE OF ASSEMBLY OF THE OMPF PORIN IN THE ENVELOPE OF ESCHERICHIA-COLI

被引:7
作者
BOLLA, JM
BERNADAC, A
LAZDUNSKI, C
PAGES, JM
机构
[1] Centre de Biochimie et de Biologie Moléculaire, CNRS, 13402 Marseille Cedex 9, 31 chemin Joseph-Aiguier
关键词
colicin receptor; export; immunolocalization; outer membrane;
D O I
10.1016/0300-9084(90)90062-L
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
Inducible hybrid genes encoding two large domains, a periplasmic domain consisting of the PhoS sequence and an outer membrane domain corresponding to various lengths of the OmpF mature sequence were constructed. The synthetisized hybrid polypeptides are correctly processed during the early times of induction, their precursor forms being accumulated at later times. These hybrids restore sensitivity toward colicin A to ompF E coli B strain which suggests an outer membrane location. At least 2 of them are indeed localized in the outer membrane after immunogold labelling on ultrathin cryosections. Insertation of a hydrophobic sequence between PhoS and OmpF improves the trimerization and the assembly of the OmpF part. Only th hybrids presenting the last C-terminal 29 residues of OmpF are able to promote the colicin N killing action and to exhibit a trimeric conformation which is recognized by specific antibodies. Moreover, the deletion of the C-terminal region impairs the functional insertion of the OmpF domain; this indicates that the last membrane-spanning region of OmpF is necessary for the correct folding and orientation of the protein in the outer membrane. © 1990.
引用
收藏
页码:385 / 395
页数:11
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