Desorption/ionization on silicon (DIOS): A diverse mass spectrometry platform for protein characterization

被引:160
作者
Thomas, JJ
Shen, ZX
Crowell, JE
Finn, MG
Siuzdak, G
机构
[1] Scripps Res Inst, Res Inst, Dept Chem, La Jolla, CA 92037 USA
[2] Scripps Res Inst, Res Inst, Dept Mol Biol, La Jolla, CA 92037 USA
[3] Scripps Res Inst, Res Inst, Skaggs Inst Chem Biol, La Jolla, CA 92037 USA
[4] Mass Consortium Corp, San Diego, CA 92121 USA
[5] Univ Calif San Diego, Dept Chem & Biochem, La Jolla, CA 92093 USA
关键词
D O I
10.1073/pnas.081069298
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Since the advent of matrix-assisted laser desorption/ionization and electrospray ionization, mass spectrometry has played an increasingly important role in protein functional characterization, increasingly important role in protein functional characterization. identification, and structural analysis. Expanding this role, desorption/ionization on silicon (DIOS) is a new approach that allows for the analysis of proteins and related small molecules, Despite the absence of matrix, DIGS-MS yields little or no fragmentation and is relatively tolerant of moderate amounts of contaminants commonly found in biological samples. Here, functional assays were performed on an esterase, a glycosidase, a lipase, as well as exo- and endoproteases by using enzyme-specific substrates. Enzyme activity also was monitored in the presence of inhibitors, successfully demonstrating the ability of DIGS to be used as an inhibitor screen, Because DIGS is a matrix-free desorption technique, it also can be used as a platform for multiple analyses to be performed on the same protein. This unique advantage was demonstrated with acetylcholine esterase for qualitative and quantitative characterization and also by its subsequent identification directly from the DIGS platform.
引用
收藏
页码:4932 / 4937
页数:6
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