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Originally published In Press as doi:10.1074/mcp.M300137-MCP200 on February 26, 2004.
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Molecular & Cellular Proteomics 3:586-595, 2004.
© 2004 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

Isolation of Outer Membrane of Synechocystis sp. PCC 6803 and Its Proteomic Characterization*

Fang Huang{ddagger},§, Erik Hedman,§, Christiane Funk||, Thomas Kieselbach, Wolfgang P. Schröder|| and Birgitta Norling{ddagger},**

From the {ddagger} Department of Biochemistry and Biophysics, Arrhenius Laboratories of Natural Sciences, Stockholm University, SE-10691 Stockholm, Sweden; Department of Medical Nutrition and Biosciences, Karolinska Institute, Novum, SE-14186 Huddinge, Sweden; and || Department of Biochemistry, Umeå University, SE-90187 Umeå, Sweden

In this report, we describe a newly developed method for isolating outer membranes from Synechocystis sp. PCC 6803 cells. The purity of the outer membrane fraction was verified by immunoblot analysis using antibodies against membrane-specific marker proteins. We investigated the protein composition of the outer membrane using two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry followed by database identification. Forty-nine proteins were identified corresponding to 29 different gene products. All of the identified proteins have a putative N-terminal signal peptide. About 40% of the proteins identified represent hypothetical proteins with unknown function. Among the proteins identified are a Toc75 homologue, a protein that was initially found in the outer envelope of chloroplasts in pea, as well as TolC, putative porins, and a pilus protein. Other proteins identified include ABC transporters and GumB, which has a suggested function in carbohydrate export. A number of proteases such as HtrA were also found in the outer membrane of Synechocystis sp. PCC 6803.

** To whom correspondence should be addressed: Department of Biochemistry and Biophysics, Arrhenius Laboratories of Natural Sciences, Stockholm University, SE-10691 Stockholm, Sweden. Tel.: 46-8-162460; Fax: 46-8-153679; E-mail: birgitta{at}dbb.su.se


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