Immobilized metal affinity chromatography and human serum proteomics

Fengrong Wang, Christyne Chmil, Frank Pierce, Kulothungan Ganapathy, Brooks B. Gump, James A. MacKenzie, Yehia Metchref, Kestutis Bendinskas

Research output: Contribution to journalArticlepeer-review

40 Scopus citations

Abstract

Metal-binding proteins have a pivotal role in normal and diseased states. We used metal affinity chromatography to enrich a fraction of human serum proteins on immobilized columns loaded with cadmium, nickel, zinc, copper, or lead in bis-Tris saline and these proteins were identified using LC-MS/MS. Tens of enriched proteins were identified and we here present the 20 most abundant for binding each metal. The binding of various proteins (complement C3, alpha-2-macroglobulin, serum albumin, apolipoprotein B-100, complement component 4B preproprotein, apolipoprotein A-I, serotransferrin, alpha-1-antitrypsin, ceruloplasmin, 47. kDa protein, uncharacterized protein DKFZp686P15220, transthyretin, hemopexin, inter-alpha-trypsin inhibitor heavy chain H2, and histidine-rich glycoprotein) to different metals using immobilized metal affinity chromatography was compared to the literature. Although many metal-binding properties of these proteins have been confirmed, new metal-binding proteins have also been identified. The metal array use in the proteomic biomarker search technologies gives this data particular importance.

Original languageEnglish (US)
Pages (from-to)26-33
Number of pages8
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume934
DOIs
StatePublished - Sep 1 2013

Keywords

  • Human serum
  • ICP-MS
  • Immobilized metal affinity chromatography
  • LC-MS/MS
  • Metal binding proteins
  • Proteomics

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology

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