title: Immobilized metal affinity chromatography and human serum proteomics.
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, 47kDa 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.
authors.affiliation: Department of Chemistry, SUNY-Oswego, Oswego, NY 13126, USA. fewang@jhsph.edu
authors.forename: Fengrong
authors.initials: F
authors.lastname: Wang
authors.forename: Christyne
authors.initials: C
authors.lastname: Chmil
authors.forename: Frank
authors.initials: F
authors.lastname: Pierce
authors.forename: Kulothungan
authors.initials: K
authors.lastname: Ganapathy
authors.forename: Brooks B
authors.initials: BB
authors.lastname: Gump
authors.forename: James A
authors.initials: JA
authors.lastname: MacKenzie
authors.forename: Yehia
authors.initials: Y
authors.lastname: Metchref
authors.forename: Kestutis
authors.initials: K
authors.lastname: Bendinskas
mesh_terms: D001798:Blood Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D008670:Metals / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D044967:Serum / Q000737:chemistry*; D053719:Tandem Mass Spectrometry
chemical_list: D001798:Blood Proteins; D008670:Metals
keywords: Human serum; ICP-MS; IDA; IMAC; Immobilized metal affinity chromatography; LC–MS/MS; Metal binding proteins; Proteomics; iminodiacetic acid; immobilized metal affinity chromatography; inductively coupled plasma mass spectrometry; liquid chromatography–tandem mass spectrometry