Phosphopeptide mapping of proteins ectopically expressed in tissue culture cell lines

Beth A. Firulli, David M. Virshup, Anthony B. Firulli

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

Post-translational modifications such as phosphorylation play a vital role in the regulation of protein function. In our study of the basic Helix-loop-Helix (bHLH) transcription factor HAND1, it was suspected that HAND1 was being phosphorylated during trophoblast giant cell differentiation and that coexpression of a constitutively active kinase with HAND1 resulted in changes in the proteins dimerization profile. In order to accurately document HAND1 phosphorylation and identify the resides being modified, we employed metabolic cell labeling with 32P of tissue culture cells coexpressing a Flag-epitope tagged HAND1 along with a number of active kinases and phosphatase subunits. We generated phosphopeptide maps of the phosphorylated HAND1 using the methods described below and linked these modifications to changes in HAND1 biological function.

Original languageEnglish (US)
Pages (from-to)16-22
Number of pages7
JournalBiological Procedures Online
Volume6
Issue number1
DOIs
StatePublished - Jan 15 2004

Keywords

  • Dimerization
  • Helix-loop-Helix mofits
  • Phosphorylation

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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