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Two‐dimensional benzyldimethyl‐ n ‐hexadecylammonium chloride/SDS‐PAGE for membrane proteomics
Author(s) -
Zahedi RenéPeiman,
Meisinger Chris,
Sickmann Albert
Publication year - 2005
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200401214
Subject(s) - proteomics , chemistry , chromatography , biochemistry , gene
Despite the importance of membranes in any living system, the global analysis of membrane subproteomes is still a common obstacle. In particular, the widely used 2‐DE technique consisting of IEF in the first dimension and SDS‐PAGE in the second dimension has some major drawbacks regarding the separation of hydrophobic proteins. Therefore, we applied an alternative electrophoretic technique for separating membrane proteins: two‐dimensional BAC/SDS electrophoresis (2‐DB) using the cationic detergent benzyldimethyl‐ n ‐hexadecylammonium chloride in the first and the anionic detergent SDS in the second dimension. The use of 2‐DB resulted in an improved separation of hydrophobic proteins. Thus, extremely hydrophobic proteins such as cytochrome‐ c oxidase subunit I with a grand average hydrophobicity (GRAVY) index of 0.74 and a total of 12 known transmembrane domains (TMD) or Sec61α with a GRAVY index of 0.56 and a total of ten known TMD could be identified by MS/MS analyses of protein spots derived from 2‐DB gels.