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Profiling of Caenorhabditis elegans proteins using two‐dimensional gel electrophoresis and matrix assisted laser desorption/ionization‐time of flight‐mass spectrometry
Author(s) -
Kaji Hiroyuki,
Tsuji Takahisa,
Mawuenyega Kwasi G.,
Wakamiya Akiko,
Taoka Masato,
Isobe Toshiaki
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(20000501)21:9<1755::aid-elps1755>3.0.co;2-s
Subject(s) - proteome , peptide mass fingerprinting , mass spectrometry , chemistry , coomassie brilliant blue , two dimensional gel electrophoresis , gel electrophoresis , matrix assisted laser desorption/ionization , polyacrylamide gel electrophoresis , time of flight mass spectrometry , protein mass spectrometry , proteomics , electrophoresis , chromatography , microbiology and biotechnology , biology , electrospray ionization , biochemistry , staining , ionization , desorption , genetics , gene , ion , organic chemistry , adsorption , enzyme
The nematode Caenorhabditis elegans ( C. elegans ) is the first animal whose whole 97 Mb genome sequence, encoding ca. 19 000 open reading frames (ORF's), has been essentially determined. We tried to establish a 2‐DE map of the nematode proteome by means of two‐dimensional polyacrylamide gel electrophoresis (2‐D PAGE). A soluble protein fraction of mixed stages of the worm, wild‐type strain N2, was applied to 2‐D PAGE. After Coomassie Brilliant Blue (CBB) staining, 1200 spots were detected and 140 major spots were excised from the gel and subjected to in‐gel digestion with Achromobacter protease I (lysyl endopeptidase). Resulting peptides were analyzed by matrix assisted laser desorption/ionization‐time of flight‐mass spectrometry (MALDI‐TOF‐MS) followed by peptide mass fingerprinting for protein identification. With this approach we have obtained a two‐dimensional electrophoresis (2‐DE) protein map in which 69 spots were localized as landmarks for comparison of expression profiles to elucidate the basis of various biological events.