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Membrane-specific targeting of green fluorescent protein by the Tat pathway in the cyanobacterium Synechocystis PCC6803

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UNSPECIFIED. (2003) Membrane-specific targeting of green fluorescent protein by the Tat pathway in the cyanobacterium Synechocystis PCC6803. MOLECULAR MICROBIOLOGY, 48 (6). pp. 1481-1489. ISSN 0950-382X

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Official URL: http://dx.doi.org/10.1046/j.1365-2958.2003.03519.x

Abstract

The transport and sorting of extracytoplasmic proteins in cyanobacteria is made complex by the presence of a highly differentiated membrane system. Proteins destined for the periplasm and thylakoid lumen are initially transported by Sec- and Tat-type pathways but little is known of the mechanisms that ultimately direct them to the correct destinations. We have generated a Synechocystis PCC6803 transformant that expresses a fusion protein comprising the Tat-specific targeting signal of Escherichia coli TorA linked to green fluorescent protein (GFP). Immunoblotting indicates the presence of mature-size GFP but no precursor form, demonstrating that efficient translocation has taken place. Confocal microscopy and immunogold electron microscopy reveal GFP to be almost exclusively located in the periplasm, with almost no protein evident in the thylakoid network. These data point to the operation of highly effective sorting pathways for soluble proteins in this cyanobacterium. The observed sorting of the GFP suggests that either (a) the Tat apparatus is located only in the plasma membrane or (b) the TorA-GFP is targeted across either membrane but the GFP is subsequently directed to the periplasm, perhaps by a default sorting pathway to this compartment.

Item Type: Journal Article
Subjects: Q Science > QD Chemistry
Q Science > QR Microbiology
Journal or Publication Title: MOLECULAR MICROBIOLOGY
Publisher: BLACKWELL PUBLISHING LTD
ISSN: 0950-382X
Date: June 2003
Volume: 48
Number: 6
Number of Pages: 9
Page Range: pp. 1481-1489
Identification Number: 10.1046/j.1365-2958.2003.03519.x
Publication Status: Published
URI: http://wrap.warwick.ac.uk/id/eprint/9661

Data sourced from Thomson Reuters' Web of Knowledge

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