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PMID: 320175 Published · ppublish English Journal Article

Mutants of Escherichia coli "cryptic" for certain periplasmic enzymes: evidence for an alteration of the outer membrane.

Journal of bacteriology ·Vol. 129 ·No. 2 ·1977-02-00 ·Pages 1034-44

Beacham IR, Haas D, Yagil E

Abstract

Mutants in which the expression of periplasmic enzymes by whole cells is reduced (termed "cryptic") are also found to show greatly reduced uptake of labeled adenosine 5'-monophosphate (5'-AMP), providing a rapid assay for crypticity. The crypticity of 3'- and 5'-nucleotidase has been examined as a function of substrate concentration. The Km for 3'- or 5'-AMP increases in the cryptic mutants when whole cells are used as the enzyme source. The Vmax is not altered. Electrophoretic analysis of protein prepared from cell envelopes showed that three cryptic mutants have a polypeptide absent from the outer membrane and a relatively high proportion of a polypeptide in the inner membrane. Analysis of the molar ratios of constituent sugars of the lipopolysaccharides showed no differences between three cryptic mutants and the parent strain. One cryptic mutant (3--41), however, has altered sensitivity to phage T4. By selection for phage resistance, derivatives of the cryptic mutants that are deoxycholate sensitive have been obtained. These mutants are no longer cryptic. We suggest that cryptic mutants have an altered outer membrane, with decreased permeability to 3'- and 5'-AMP, as a result of an altered polypeptide.

MeSH Terms
Adenosine Monophosphate/metabolism Bacterial Proteins/analysis Cell Wall/metabolism,ultrastructure Escherichia coli/enzymology,metabolism,ultrastructure Galactose/analysis Glucose/analysis Heptoses/analysis Ketoses Lipopolysaccharides/analysis Mutation Nucleotidases/metabolism Peptides/analysis Permeability Phosphates/analysis Polysaccharides, Bacterial/analysis Sugar Acids/analysis
Chemicals
Bacterial Proteins Heptoses Ketoses Lipopolysaccharides Peptides Phosphates Polysaccharides, Bacterial Sugar Acids Adenosine Monophosphate Nucleotidases Glucose Galactose
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Beacham I R
Haas D
Yagil E
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43 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.129.2.1034-1044.1977
Published
1977-02-00
Pages
1034-44
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC235043
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.129.2.1034-1044.1977 PMC Full Text View journal details
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