Journal article
Interaction of the Gonococcal Porin P.IB with G- and F-Actin
Biochemistry (Easton), Vol.39(29), pp.8638-8647
07/25/2000
DOI: 10.1021/bi000241j
PMID: 10913272
Abstract
The invasion of epithelial cells by N. gonorrheae is accompanied by formation of a halo of actin filaments around the enveloped bacterium. The transfer of the bacterial major outer membrane protein, porin, to the host cell membrane during invasion makes it a candidate for a facilitator for the formation of this halo. Western analysis shows here that gonococcal porin P.IB associates with the actin cytoskeleton in infected cells. Using the pyrene-labeled Mg forms of yeast and muscle actins, we demonstrate that under low ionic strength conditions, P.IB causes formation of filamentous actin assemblies, although they, unlike F-actin, cannot be internally cross-linked with N,N‘-4-phenylenedimaleimide (PDM). In F-buffer, low porin concentrations appear to accelerate actin polymerization. Higher P.IB concentrations lead to the formation of highly decorated fragmented F-actin-like filaments in which the actin can be cross-linked by PDM. Co-assembly of P.IB with a pyrene-labeled mutant actin, S265C, prevents formation of a pyrene excimer present with labeled S265C F-actin alone. Addition of low concentrations of porin to preformed F-actin results in sparsely decorated F-actin. Higher P.IB concentrations extensively decorate the filaments, thereby altering their morphology to a state like that observed when the components are copolymerized. With preformed labeled S265C F-actin, P.IB quenches the pyrene excimer. This decrease is prevented by the F-actin stabilizers phalloidin and to a lesser extent beryllium fluoride. P.IB's association with the actin cytoskeleton and its ability to interact with and remodel actin filaments support a direct role for porin in altering the host cell cytoskeleton during invasion.
Details
- Title: Subtitle
- Interaction of the Gonococcal Porin P.IB with G- and F-Actin
- Creators
- Kuo-Kuang WenPeter C GiardinaMilan S BlakeJennifer EdwardsMichael A ApicellaPeter A Rubenstein
- Resource Type
- Journal article
- Publication Details
- Biochemistry (Easton), Vol.39(29), pp.8638-8647
- Publisher
- American Chemical Society
- DOI
- 10.1021/bi000241j
- PMID
- 10913272
- ISSN
- 0006-2960
- eISSN
- 1520-4995
- Language
- English
- Date published
- 07/25/2000
- Academic Unit
- Microbiology and Immunology; Stead Family Department of Pediatrics; Biochemistry and Molecular Biology; Internal Medicine
- Record Identifier
- 9984025255302771
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