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.
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 gel electrophoresis. Light-induced protein spots electroblotted on a polyvinyldiene difluoride membrane were analyzed by N-terminal Edman sequence determination and followed by CyanoBase. The tryptic 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.
Choi
J S
JS
Biomolecule Research Team, Korea Basic Science Institute, Taejon, Korea. jschoi@comp.kbsi.re.kr
Kim
D S
DS
Lee
J
J
Kim
S J
SJ
Kim
S I
SI
Kim
Y H
YH
Hong
J
J
Yoo
J S
JS
Suh
K H
KH
Park
Y M
YM
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis; D000458:Cyanobacteria / Q000737:chemistry* / Q000528:radiation effects; D015180:Electrophoresis, Gel, Two-Dimensional; D015870:Gene Expression / Q000528:radiation effects*; D020869:Gene Expression Profiling; D008027:Light; D008969:Molecular Sequence Data; D009943:Organophosphorus Compounds; D020543:Proteome / Q000032:analysis* / Q000528:radiation effects; D020539:Sequence Analysis, Protein; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
C043517:2-(4-isothiocyanatophenoxy)-1,3,2-dioxaphosphinene 2-oxide; D001426:Bacterial Proteins; D009943:Organophosphorus Compounds; D020543:Proteome