TRANSLOCATION OF A FOLDED PROTEIN ACROSS THE OUTER-MEMBRANE IN ESCHERICHIA-COLI

被引:98
作者
PUGSLEY, AP
机构
[1] Unite de Genetique Moleculaire, CNRS, Institut Pasteur, Paris 75724 Cedex 15, 25, rue du Dr. Roux
关键词
PROTEIN SECRETION; PULLULANASE; DISULFIDE BOND;
D O I
10.1073/pnas.89.24.12058
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
A mutation in the Escherichia coli dsbA gene (coding for a periplasmic disulfide oxidoreductase) reduces the rate of disulfide bond formation in the enzyme pullulanase and also reduces the rate at which the enzyme is secreted to the cell surface, as measured by protease accessibility. The enzyme did not become protease accessible when disulfide bond formation was completely prevented in the mutant strain by carboxymethylation. These results indicate that a disulfide bond may be required for, and certainly does not impede, the translocation of pullulanase across the outer membrane. Since it is unlikely that a disulfide bond could be formed and then reduced again in the periplasm, these results would appear to strengthen the argument that pullulanase polypeptides fold into or close to their final conformation before they are transported across the outer membrane. It is suggested that this might be a feature common to all proteins that are secreted by other Gram-negative bacteria by a pullulanase-like pathway.
引用
收藏
页码:12058 / 12062
页数:5
相关论文
共 28 条
[1]   IDENTIFICATION OF A PROTEIN REQUIRED FOR DISULFIDE BOND FORMATION INVIVO [J].
BARDWELL, JCA ;
MCGOVERN, K ;
BECKWITH, J .
CELL, 1991, 67 (03) :581-589
[2]   SECRETION AND MEMBRANE INTEGRATION OF A FILAMENTOUS PHAGE-ENCODED MORPHOGENETIC PROTEIN [J].
BRISSETTE, JL ;
RUSSEL, M .
JOURNAL OF MOLECULAR BIOLOGY, 1990, 211 (03) :565-580
[3]   DOMINANT CONSTITUTIVE MUTATIONS IN MALT, POSITIVE REGULATOR GENE OF MALTOSE REGULON IN ESCHERICHIA-COLI [J].
DEBARBOUILLE, M ;
SHUMAN, HA ;
SILHAVY, TJ ;
SCHWARTZ, M .
JOURNAL OF MOLECULAR BIOLOGY, 1978, 124 (02) :359-371
[4]  
DENFERT C, 1989, J BIOL CHEM, V264, P17462
[5]   CLONING AND EXPRESSION IN ESCHERICHIA-COLI OF THE KLEBSIELLA-PNEUMONIAE GENES FOR PRODUCTION, SURFACE LOCALIZATION AND SECRETION OF THE LIPOPROTEIN PULLULANASE [J].
DENFERT, C ;
RYTER, A ;
PUGSLEY, AP .
EMBO JOURNAL, 1987, 6 (11) :3531-3538
[6]   CONFORMATION OF PROTEIN SECRETED ACROSS BACTERIAL OUTER MEMBRANES - A STUDY OF ENTEROTOXIN TRANSLOCATION FROM VIBRIO-CHOLERAE [J].
HIRST, TR ;
HOLMGREN, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1987, 84 (21) :7418-7422
[7]   IDENTIFICATION AND CHARACTERIZATION OF AN ESCHERICHIA-COLI GENE REQUIRED FOR THE FORMATION OF CORRECTLY FOLDED ALKALINE-PHOSPHATASE, A PERIPLASMIC ENZYME [J].
KAMITANI, S ;
AKIYAMA, Y ;
ITO, K .
EMBO JOURNAL, 1992, 11 (01) :57-62
[8]   MOLECULAR CHARACTERIZATION OF PULA AND ITS PRODUCT, PULLULANASE, A SECRETED ENZYME OF KLEBSIELLA-PNEUMONIAE UNF5023 [J].
KORNACKER, MG ;
PUGSLEY, AP .
MOLECULAR MICROBIOLOGY, 1990, 4 (01) :73-85
[9]   THE NORMALLY PERIPLASMIC ENZYME BETA-LACTAMASE IS SPECIFICALLY AND EFFICIENTLY TRANSLOCATED THROUGH THE ESCHERICHIA-COLI OUTER-MEMBRANE WHEN IT IS FUSED TO THE CELL-SURFACE ENZYME PULLULANASE [J].
KORNACKER, MG ;
PUGSLEY, AP .
MOLECULAR MICROBIOLOGY, 1990, 4 (07) :1101-1109
[10]   MOLECULAR CHAPERONES AND PROTEIN TRANSLOCATION ACROSS THE ESCHERICHIA-COLI INNER MEMBRANE [J].
KUMAMOTO, CA .
MOLECULAR MICROBIOLOGY, 1991, 5 (01) :19-22