Evidence of multimeric forms of HSP70 with phosphorylation on serine and tyrosine residues - implications for roles of HSP70 in detection of gi cancers

Anand Dutta, Mohit Girotra, Nipun Merchant, Padmanabhan Nair, Sudhir Kumar Dutta

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Background: Heat-shock protein70 (HSP70) are intracellular protein chaperones, with emerging evidence of their association with various diseases. We have previously reported significantly elevated plasma-HSP70 (pHSP70) in pancreatic cancer. Current methods of pHSP70 isolation are ELISA-based which lack specificity due to cross-reactivity by similarities in the amino-acid sequence in regions of the protein backbone resulting in overestimated HSP70 value. Materials and Methods: This study was undertaken to develop a methodology to capture all isoforms of pHSP70, while further defining their tyrosine and serine phosphorylation status. Results: The methodology included gel electrophoresis on centrifuged supernatant obtained from plasma incubated with HSP70 antibody-coupled beads. After blocking non-specific binding sites, blots were immunostained with monoclonal-antibody specific for human-HSP70, phosphoserine and phosphotyrosine. Conclusions: Our novel immunocapture approach has distinct advantages over the commercially available methods of pHSP70 quantification by allowing isolation of molecular aggregates of HSP70 with additional ability to precisely distinguish phosphorylation state of HSP70 molecules at serine and tyrosine residues.

Original languageEnglish (US)
Pages (from-to)5741-5745
Number of pages5
JournalAsian Pacific Journal of Cancer Prevention
Volume14
Issue number10
DOIs
StatePublished - 2013

Keywords

  • GI cancer detection
  • HSP70
  • Multimeric forms
  • Phosphorylation
  • Serine
  • Tyrosine

ASJC Scopus subject areas

  • Epidemiology
  • Oncology
  • Public Health, Environmental and Occupational Health
  • Cancer Research

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