1887

Abstract

The previous discovery of the gene and its overexpression followed by the functional reconstitution of the purified gene product has resulted in new strategies to explore this channel protein . KcsA has evolved as a general model to investigate the structure/function relationship of ion channel proteins. Using specific antibodies raised against a domain of KcsA lacking membrane-spanning regions, KcsA has now been localized within numerous separated clusters between the outer face of the cytoplasm and the cell envelope in substrate hyphae of the wild-type strain but not in a designed chromosomal disruption mutant ΔK, lacking a functional gene. Previous findings had revealed that caesium ions led to a block of KcsA channel activity within protoplasts fused to giant vesicles. As caesium can be scored by electron energy loss spectroscopy better than potassium, this technique was applied to hyphae that had been briefly exposed to caesium instead of potassium ions. Caesium was found preferentially at the cell envelope. Compared to the ΔK mutant, the relative level of caesium was ≈30 % enhanced in the wild-type. This is attributed to the presence of KcsA channels. Additional visualization by electron spectroscopic imaging supported this conclusion. The data presented are believed to represent the first demonstration of monitoring of KcsA in its original host.

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2006-09-01
2020-04-01
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