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PMID: 3025168 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Isolation of the outer membrane and characterization of the major outer membrane protein from Spirochaeta aurantia.

Journal of bacteriology ·Vol. 169 ·No. 1 ·1987-01-00 ·Pages 172-9

Kropinski AM, Parr TR, Angus BL, Hancock RE, Ghiorse WC, Greenberg EP

Abstract

The outer membrane of Spirochaeta aurantia was isolated after cells were extracted with sodium lauryl sarcosinate and was subsequently purified by differential centrifugation and KBr isopycnic gradient centrifugation. The purified outer membrane was obtained in the form of carotenoid-containing vesicles. Four protein species with apparent molecular weights of 26,000 (26K), 36.5K, 41K, and 48.5K were readily observed as components of the vesicles. The 36.5K protein was the major polypeptide and constituted approximately 90% of the outer membrane protein observed on sodium dodecyl sulfate-polyacrylamide gels. Under mild denaturing conditions the 36.5K major protein exhibited an apparent molecular weight of approximately 90,000. This, together with the results of protein cross-linking studies, indicates that the 36.5K polypeptide has an oligomeric conformation in the native state. Reconstitution of solubilized S. aurantia outer membrane into lipid bilayer membranes revealed the presence of a porin, presumably the 36.5K protein, with an estimated channel diameter of 2.3 nm based on the measured single channel conductance of 7.7 nS in 1 M KCl.

MeSH Terms
Bacterial Outer Membrane Proteins/analysis,isolation & purification Electrophoresis, Polyacrylamide Gel Hot Temperature Microscopy, Electron Molecular Weight Porins Spirochaeta/analysis
Chemicals
Bacterial Outer Membrane Proteins Porins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kropinski A M
Parr T R
Angus B L
Hancock R E
Ghiorse W C
Greenberg E P
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.169.1.172-179.1987
Published
1987-01-00
Pages
172-9
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC211749
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.169.1.172-179.1987 PMC Full Text View journal details
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