Premium
Amino acid sequence of the signal peptide of OmpF, a major outer membrane protein of Escherichia coli
Author(s) -
Mutoh Norihiro,
Inokuchi Kaoru,
Mizushima Shoji
Publication year - 1982
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(82)80341-4
Subject(s) - library science , biological sciences , escherichia coli , agriculture , microbiology and biotechnology , chemistry , biology , computer science , biochemistry , ecology , gene
Studies on a wide variety of secretory proteins have substantiated the signal hypothesis [ 11. The signal hypothesis states that a protein exported through a membrane is synthesized as a precursor form with a peptide extension at the NHz-terminal end (signal peptide). The cell envelope of Gram-negative bacteria consists of 2 membranous organelles; the outer membrane and the cytoplasmic membrane. Therefore, outer membrane proteins that are synthesized in the cytoplasm have to be exported through the cytoplasmic membrane. Indeed, almost all of the outer membrane proteins so far studied were found to be first synthesized as a precursor form, and in 3 cases (OmpA [2], major lipoprotein [3] and LamB [4]) the amino acid sequence of the signal peptide has been determined. The OmpF protein is another major outer membrane protein to be studied. To study the chemical structure of OmpF and its precursor as well as the mechanism controlling the synthesis and subsequent localization in the outer membrane of this protein, we isolated a specialized transducing X phage that carries the ompF gene [S]. Here, we have subcloned the ompF gene and determined the DNA sequence that covers the NHz-terminal region of OmpF. From the DNA sequence the amino acid sequence of the signal peptide was deduced. The general features of the signal peptides and those of the cleavage site are discussed in relation to protein localization in the cell surface. and ADP (Sigma Chemicals) as m [6], and used for DNA sequencing. Restriction endonucleases EcoRI, HaeIII, HapII, HindIII, HhaI and Sal1 were products of Takara Shuzo Co. and PstI, PvuII and TaqI were from New England Biolabs. Bacterial alkaline phosphatase was from Worthington Biochemical Corp., bacteriophage T4 polynucleotide kinase from P. L. Biochemicals, and T4 ligase from Takara Shuzo Co.