Novel techniques for identification and characterization of proteins loaded on gels in femtomole amounts

被引：16

作者：

Gevaert, K

DeMol, H

Verschelde, JL

VanDamme, J

DeBoeck, S

Vandekerckhove, J

机构：

[1] Universiteit Gent,Flanders Interuniversity Institute for Biotechnology, Department of Biochemistry, Faculty of Medicine

来源：

JOURNAL OF PROTEIN CHEMISTRY | 1997年 / 16卷 / 05期

关键词：

gel electrophoresis; peptide femtomole sequencing; peptide mass fingerprinting; matrix-assisted-laser-desorption-ionization; post source decay;

D O I：

10.1023/A:1026320318489

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A combination of techniques is presented allowing gel-purified protein identification in the femtomole range using matrix-assisted-laser-desorption-ionization mass spectrometry. The proteins are detected in the primary gel by a sensitive negative staining procedure, transferred, and concentrated in a secondary gel matrix, There, they are digested in the presence of (H2O)-O-18 and their sequences are predicted (1) by peptide mass fingerprinting, (2) by comparing the post-source-decay (PSD) spectra with theoretical spectra of candidate isobaric peptides using a computer algorithm called MassFrag, and (3) by a manual readout of the O-18/O-16-labeled fragmentation ions in the PSD spectra.

引用

页码：335 / 342

页数：8

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