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PMID: 1708384 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Membrane topography of ColE1 gene products: the immunity protein.

Journal of bacteriology ·Vol. 173 ·No. 9 ·1991-05-00 ·Pages 2935-43

Song HY, Cramer WA

Abstract

The topography of the colicin E1 immunity (Imm) protein was determined from the positions of TnphoA and complementary lacZ fusions relative to the three long hydrophobic segments of the protein and site-directed substitution of charged for nonpolar residues in the proposed membrane-spanning segments. Inactivation of the Imm protein function required substitution and insertion of two such charges. It was concluded that the 113-residue colicin E1 Imm protein folds in the membrane as three trans-membrane alpha-helices, with the NH2 and COOH termini on the cytoplasmic and periplasmic sides of the membrane, respectively. The approximate spans of the three helices are Asn-9 to Ser-28, Ile-43 to Phe-62, and Leu-84 to Leu-104. An extrinsic highly charged segment, Lys-66 to Lys-74, containing seven charges in nine residues, extends into the cytoplasmic domain. The specificity of the colicin E1 Imm protein for interaction with the translocation apparatus and the colicin E1 ion channel is proposed to reside in its peripheral segments exposed on the surface of the inner membrane. These regions include the highly charged segment Lys-66 to Lys-83 (loop 2) and the short (approximately eight-residue) NH2 terminus on the cytoplasmic side, and Glu-29 to Val-44 (loop 1) and the COOH-terminal segment Gly-105 to Asn-113 on the periplasmic side.

Related Genes
TmphoA lacZ
MeSH Terms
Alkaline Phosphatase/metabolism Amino Acid Sequence Bacterial Proteins/biosynthesis,chemistry Biological Transport Cell Membrane/metabolism,ultrastructure Cloning, Molecular Colicins/antagonists & inhibitors Ion Channels Models, Biological Molecular Sequence Data Mutagenesis, Insertional Mutagenesis, Site-Directed Plasmids Sequence Homology, Nucleic Acid beta-Galactosidase/metabolism
Chemicals
Bacterial Proteins Colicins Ion Channels colicin immunity proteins Alkaline Phosphatase beta-Galactosidase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Song H Y
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907.
Cramer W A
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41 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.173.9.2935-2943.1991
Published
1991-05-00
Pages
2935-43
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC207876
Subset
IM
Grants
NIGMS NIH HHS · GM-18457 · United States
External Links
PubMed source DOI: 10.1128/jb.173.9.2935-2943.1991 PMC Full Text View journal details
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