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Methods. 2008 Feb;44(2):164-9. doi: 10.1016/j.ymeth.2007.09.002.

Monitoring Lys-tRNA(Lys) phosphatidylglycerol transferase activity.

Author information

1
Department of Microbiology, The Ohio State University, 484 West 12th Avenue, Columbus, OH 43210, USA. roy.79@osu.edu

Abstract

In some bacteria Lys-tRNA(Lys) is used both in translation and for the specific addition of Lys to phosphatidylglycerol in the cytoplasmic membrane. This reaction is catalyzed by the membrane protein MprF, and the lysyl-phosphatidylglycerol formed contributes to the resistance of these bacteria to various cationic antibacterial molecules. Obtaining proteins and reconstituting an in vitro system mimicking membrane conditions is a major challenge to studying the function of membrane proteins, especially when labile substrates such as Lys-tRNA(Lys) are required. Here we report methods to obtain a stable enriched membrane fraction containing MprF, and the techniques necessary to quantitatively monitor its activity in vitro and in vivo.

PMID:
18241797
PMCID:
PMC2271063
DOI:
10.1016/j.ymeth.2007.09.002
[Indexed for MEDLINE]
Free PMC Article

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