51. Isolation of outer membrane of Synechocystis sp. PCC 6803 and its proteomic characterization.
- Author
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Huang F, Hedman E, Funk C, Kieselbach T, Schröder WP, and Norling B
- Subjects
- Amino Acid Sequence, Bacterial Proteins isolation & purification, Cell Membrane metabolism, Electrophoresis, Gel, Two-Dimensional methods, Molecular Sequence Data, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization methods, Subcellular Fractions metabolism, Bacterial Outer Membrane Proteins analysis, Bacterial Proteins analysis, Cell Membrane chemistry, Proteome analysis, Synechocystis chemistry
- Abstract
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.
- Published
- 2004
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