Proteomics

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Improved protease digestion conditions for membrane protein detection


ABSTRACT: This work presents improved protease digestion conditions for membrane protein detection. Two improved conditions, 0.01% SDS or the combination of 10% MeOH and 0.01% RapiGest, were chosen and applied to proteins from Escherichia coli. The application of both improved conditions to a membrane protein fraction of Escherichia coli resulted in the identification of 309 (SDS) and 329 (MeOH/RapiGest) unique proteins of which 140/309 and 148/329 were membrane proteins.

INSTRUMENT(S): Q TRAP

ORGANISM(S): Escherichia Coli

SUBMITTER: Lie Min  

LAB HEAD: Kelvin H. Lee

PROVIDER: PXD001824 | Pride | 2015-04-28

REPOSITORIES: Pride

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Improved protease digestion conditions for membrane protein detection.

Min Lie L   Choe Leila H LH   Lee Kelvin H KH  

Electrophoresis 20150610 15


This work presents improved protease digestion conditions for membrane protein detection. The enzymatic digest of bacteriorhodopsin (BR), a model membrane protein with seven transmembrane domains (TMDs) was investigated. An initial in-gel digestion identified 17% BR sequence coverage, including part of the seventh TMD. To improve sequence coverage, BR digest was tested with different concentrations of RapiGest, methanol (MeOH) and SDS using either trypsin or chymotrypsin. Two improved conditions  ...[more]

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