MALDI-TOF/TOF de novo sequence analysis of 2-D PAGE-separated proteins from Halorhodospira halophila, a bacterium with unsequenced genome

被引：28

作者：

Samyn, Bart ^{[1
]}

Sergeant, Kjell ^{[1
]}

Memmi, Samy ^{[1
]}

Debyser, Griet ^{[1
]}

Devreese, Bart ^{[1
]}

Van Beeumen, Jozef ^{[1
]}

机构：

[1] Univ Ghent, Dept Biochem Physiol & Microbiol, Lab Prot Biochem & Prot Engn, B-9000 Ghent, Belgium

来源：

ELECTROPHORESIS | 2006年 / 27卷 / 13期

关键词：

chemically assisted fragmentation; guanidination; homology search; sulfonation;

D O I：

10.1002/elps.200500959

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Because protein identifications rely on matches with sequence databases, high-throughput proteomics is currently largely restricted to those species for which comprehensive sequence databases are available. The identification of proteins derived from organisms with unsequenced genomes mainly depends on homology searching. Here, we report the use of a simplified, gel-based, chemical derivatization strategy for de novo sequence analysis using a MALDI-TOF/TOF mass spectrometer. This approach allows the determination of de novo peptide sequences of up to 20 amino acid residues in length. The protocol was applied on a proteomic study of 2-D PAGE-separated proteins from Halorhodospira halophila, an extremophilic eubacterium with yet unsequenced genome. Using three different homology-based search algorithms, we were able to identify more than 30 proteins from this organism using subpicomole quantities of protein.

引用

页码：2702 / 2711

页数：10

共 29 条

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