A method for analyzing lipid-modified proteins with mass spectrometry.
In protein analysis using mass spectrometry, proteins are usually separated by electrophoresis and digested within the gel with proteases such as trypsin. However, analysis of lipid-modified proteins is difficult due to the low recovery of lipid-modified peptide fragments from the gel as well as their low ionization efficiency during mass spectrometry. In this study, we developed a simple extraction method with n-dodecyl-beta-D-maltoside following chloroform/methanol extraction that efficiently elutes lipid-modified fragments from gels. This method allowed us to analyze the structure of lipid-modified fragments, suggesting the applicability of the method for analysis of lipid-modified fragments by mass spectrometry.
Ujihara
Tetsuro
T
Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Meguro-ku, Tokyo 153-8902, Japan.
Sakurai
Isamu
I
Mizusawa
Naoki
N
Wada
Hajime
H
D000595:Amino Acid Sequence; D002725:Chloroform / Q000737:chemistry; D004586:Electrophoresis; D005960:Glucosides / Q000737:chemistry; D008055:Lipids / Q000032:analysis* / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D000432:Methanol / Q000737:chemistry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry; D014357:Trypsin / Q000737:chemistry / Q000378:metabolism
D005960:Glucosides; D008055:Lipids; D010446:Peptide Fragments; D011506:Proteins; C040358:dodecyl maltoside; D002725:Chloroform; D014357:Trypsin; D000432:Methanol