Abstract
Previous studies demonstrated that a cloned 2-megadalton (MDal) fragment of Escherichia coli DNA contained the structural gene for major outer membrane protein a (also known as 3b or M2 (40 kDal). The present study demonstrates that M2 is synthesized from a 42-kDal precursor that also is present in the outer membrane. The conversion of the 42-kDal precursor to M2 is inhibited by a number of different local anesthetics (procaine, piperocaine, lidocaine, cocaine), by the neuroactive drug atropine, and by the classical trypsin inhibitors N alpha-tosyllysine chloromethyl ketone (TLCK) and benzamidine. Our kinetic studies demonstrate that the amidase action of pure trypsin is inhibited competitively by the local anesthetics tested (excluding lidocaine) as well as by atropine and neostigmine. A mechanism of action for local anesthetics as well as atropine in E. coli may to be inhibit trypsinlike proteases, in a competitive manner, in the region of the outer membrane. The mechanism of action of these compounds in regulating nerve conduction in man have certain features in common with the mechanism proposed in E. coli.
MeSH Terms
Anesthetics, Local/pharmacology
Atropine/pharmacology
Bacterial Proteins/metabolism
Chloramphenicol/pharmacology
Escherichia coli
Membrane Fluidity
Membrane Proteins/metabolism
Plasmids
Procaine/pharmacology
Protein Precursors/metabolism
Temperature
Trypsin/metabolism
Trypsin Inhibitors/pharmacology
Chemicals
Anesthetics, Local
Bacterial Proteins
Membrane Proteins
Protein Precursors
Trypsin Inhibitors
Procaine
Chloramphenicol
Atropine
Trypsin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Gayda R C
Henderson G W
Markovitz A
References (23)
23 references, click to expand
-
STUDIES ON THE ACTIVE CENTER OF TRYPSIN. THE BINDING OF AMIDINES AND GUANIDINES AS MODELS OF THE SUBSTRATE SIDE CHAIN.
J Biol Chem. 1965 Apr;240:1579-85
PMID: 14285494
-
Statistical estimations in enzyme kinetics.
Biochem J. 1961 Aug;80:324-32
PMID: 13785321
-
Vesicle penicillinase of Bacillus licheniformis: existence of periplasmic-releasing factor(s).
J Bacteriol. 1977 Jan;129(1):184-90
PMID: 12136
-
Deg phenotype of Escherichia coli lon mutants.
J Bacteriol. 1978 Feb;133(2):844-51
PMID: 146704
-
Bacteriophage infection of minicells: a general method for identification of "in vivo" bacteriophage directed polypeptide biosynthesis.
Mol Gen Genet. 1977 Dec 14;158(1):73-9
PMID: 415222
-
Cloned DNA fragment specifying major outer membrane protein a in Escherichia coli K-12.
J Bacteriol. 1978 Oct;136(1):369-80
PMID: 361698
-
An Escherichia coli mutant with an amino acid alteration within the signal sequence of outer membrane prolipoprotein.
Proc Natl Acad Sci U S A. 1978 Oct;75(10):4891-5
PMID: 368803
-
The outer membrane proteins of Gram-negative bacteria: biosynthesis, assembly, and functions.
Annu Rev Biochem. 1978;47:481-532
PMID: 354502
-
Precursors of three exported proteins in Escherichia coli.
Proc Natl Acad Sci U S A. 1978 Mar;75(3):1209-12
PMID: 349560
-
Intermediate steps in the degradation of a specific abnormal protein in Escherichia coli.
J Biol Chem. 1977 Dec 10;252(23):8350-7
PMID: 336618
-
Outer membrane proteins of Escherichia coli. VI. Protein alteration in bacteriophage-resistant mutants.
J Bacteriol. 1977 Aug;131(2):608-22
PMID: 328489
-
Induction of alkaline phosphatase in Escherichia coli: effect of procaine hydrochloride.
J Bacteriol. 1977 Aug;131(2):431-7
PMID: 328482
-
Synthesis and processing of an Escherichia coli alkaline phosphatase precursor in vitro.
Proc Natl Acad Sci U S A. 1977 Apr;74(4):1440-4
PMID: 323853
-
Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis.
J Biol Chem. 1977 Feb 10;252(3):1102-6
PMID: 320200
-
Effects of heating in dodecyl sulfate solution on the conformation and electrophoretic mobility of isolated major outer membrane proteins from Escherichia coli K-12.
J Biochem. 1976 Dec;80(6):1411-22
PMID: 828162
-
Influence of cultural conditions and mutations on the composition of the outer membrane proteins of Escherichia coli.
Mol Gen Genet. 1976 Sep 23;147(3):251-62
PMID: 787762
-
The effect of local anaesthetic agents on lipolysis by human adipose tissue.
Life Sci. 1973 Jul 16;13(2):161-9
PMID: 4356417
-
The ion gene and degradation of beta-galactosidase nonsense fragments.
J Bacteriol. 1973 Dec;116(3):1469-71
PMID: 4584816
-
A regulator gene that is dominant on an episome and recessive on a chromosome.
Proc Natl Acad Sci U S A. 1965 Oct;54(4):1084-91
PMID: 5327253
-
Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma.
J Cell Biol. 1975 Dec;67(3):835-51
PMID: 811671
-
Cloning of Escherichia coli DNA that controls cell division and capsular polysaccharide synthesis.
Proc Natl Acad Sci U S A. 1976 Mar;73(3):697-701
PMID: 768981
-
Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands.
FEBS Lett. 1975 Oct 15;58(1):254-8
PMID: 773686
-
Local anesthetics affect transmembrane cytoskeletal control of mobility and distribution of cell surface receptors.
Proc Natl Acad Sci U S A. 1975 Nov;72(11):4430-4
PMID: 1060123