Two‐dimensional gel electrophoresis of <i>Caenorhabditis elegans</i> homogenates and identification of protein spots by microsequencing

書誌事項

公開日
1997-01
権利情報
  • http://onlinelibrary.wiley.com/termsAndConditions#vor
DOI
  • 10.1002/elps.1150180337
公開者
Wiley

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説明

<jats:title>Abstract</jats:title><jats:p>Employing isoelectric focusing on immobilized pH gradients followed by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) we have obtained a map of <jats:italic>C. elegans</jats:italic> proteins, from a mixed culture containing all developmental stages, presenting over 2000 spots within the window of isoelectric points (p<jats:italic>I</jats:italic>) 3.5–9 and a molecular mass of 10–200 kDa. Edman microsequencing yielded successful results in 12 out of 24 analyzed spots. All but one of the <jats:italic>N</jats:italic>‐terminal sequences retrieved <jats:italic>C. elegans</jats:italic> sequences in cosmid and/or expressed sequence tag clones. Structurally related protein sequences found in data banks included enzymes in energy metabolism (cytochrome oxydase, ATP synthase, enolase), a fatty acid‐binding protein, a translationally controlled tumor protein, an unknown <jats:italic>C. elegans</jats:italic> protein, an acidic ribosomal protein, a titin‐like protein, a G‐protein β chain, cyclophilin, and cathepsin D. Experimental determination of <jats:italic>N</jats:italic>‐termini allowed us to define sites of signal cleavage providing further information on the physiological role of the newly found <jats:italic>C. elegans</jats:italic> proteins. This report demonstrates the possibility of two‐dimensional gel electrophoresis and Edman microsequencing in the elucidation of <jats:italic>C. elegans</jats:italic> proteome.</jats:p>

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