Proteome analysis of light-induced proteins in Synechocystis sp PCC 6803:: Identification of proteins separated by 2D-PAGE using N-terminal sequencing and MALDI-TOF MS

被引:24
作者
Choi, JS [1 ]
Kim, DS
Lee, J
Kim, SJ
Kim, SI
Kim, YH
Hong, J
Yoo, JS
Suh, KH
Park, YM
机构
[1] Korea Basic Sci Inst, Biomol Res Team, Taejon 305333, South Korea
[2] Paichai Univ, Dept Biochem & Biomed RRc, Taejon 302735, South Korea
[3] Korea Basic Sci Inst, Mass Spectrometry Anal Team, Taejon 305333, South Korea
关键词
2D-PAGE; MALDI-TOF; mass finger printing; N-terminal sequencing;
D O I
10.1007/s100590000033
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
The cyanobacterium Synechocystis sp. PCC 6803 is an ideal model organism for the proteome study of light-induced gene expression because the whole genomic sequence has been determined. The soluble proteins extracted from light- and dark-cultured cells were separated by two-dimensional polyacrylamide get electrophoresis. Light-induced protein spots electroblotted on a polyvinyldiene difluoride membrane were analyzed by N-terminal Edman sequence determination and followed by CyanoBase. The tryptie digests of some proteins were also confirmed by matrix-assisted laser desorption ionization/time-of-flight (MALDI-TOF) and MS-Fit search. Interestingly, eight proteins were related to photosynthesis and respiration (RbcS/L, CbbA, Gap2, AtpB, CpcB, PsbO, and PsbU). Four proteins (SodB, DnaK, GroEL2, and Tig) were involved in cellular processes and the functions of another two proteins (rehydrin and membrane protein) were unknown. The proteome analysis by N-terminal Edman sequencing and MALDI-TOF enabled us to characterize one-shot protein profiles expressed under different physiological conditions.
引用
收藏
页码:705 / 711
页数:7
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