A Simple Search of TM Segments in Polytopic Membrane Protein Using Matrix - Assisted Laser DesorptionIonization Time - of - Flight Mass Spectrometry

Author(s): Yoshito Abe, Tomohiro Hamasaki, Susumu Turusaki, Shinya Takazaki, Xiuri Jin, Dongchon Kang, Naotaka Hamasaki.

Journal Name: Protein & Peptide Letters

Volume 13 , Issue 8 , 2006

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Abstract:

Using both high performance liquid chromatography (HPLC) and amino acid sequencing (AAS), we previously analyzed band 3 TM peptide-segments that make up the transmembrane protein structure. However, the HPLC/AAS combination method was highly time-consuming. Matrix-Assisted Laser Desorption Ionization Time-of-Flight (MALDI-TOF) mass spectrometry is used to obtain accurate molecular weight information for proteins/peptides simply and sensitively. We applied the MALDI-TOF mass spectrometry technique to search for TM segments in membrane proteins. In combination with trypsin cleavages after alkali treatments (pH12 or 13) and sample preparation using organic solvents for MALDI-TOF mass spectrometry, we determined the TM segments of band 3 and glycophorin A in erythrocyte membrane. The method can be applied to other polytopic membrane proteins in erythrocyte membrane.

Keywords: Transmembrane, Polytopic membrane protein, MALDI-TOF mass spectrometry

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Article Details

VOLUME: 13
ISSUE: 8
Year: 2006
Page: [761 - 767]
Pages: 7
DOI: 10.2174/092986606777841316
Price: $58