title: Evaluating immobilized metal affinity chromatography for the selection of histidine-containing peptides in comparative proteomics.
abstract: Agarose based immobilized metal affinity chromatography (IMAC) columns loaded with copper (II) were evaluated for the selection of histidine-containing peptides in comparative proteomics. Recovery, binding specificity, and reproducibility were investigated with model proteins. Cu(II)-IMAC was found to be highly selective for histidine containing peptides; moreover, a low degree of nonspecific selection was observed. Acylation of the amino-terminus of peptides with either succinic anhydride, N-acetoxysuccinamide, or [3-(2,5)-dioxopyrrolidin-1-yloxycarbonyl)-propyl]-trimethylammonium (quaternary amine) reduced the number of histidine-containing peptides bound by the Cu(II)-IMAC columns. This provides an additional possibility for sample simplification in proteomic applications. The number of acylated peptides selected decreased in the order of quaternary amine > N-acetoxysuccinamide > succinic anhydride derivatization. Although the selection of N-terminally derivatized peptides is biased toward peptides that contain more than one histidine, it is not yet possible to predict selectivity.
authors.affiliation: Department of Chemistry, Purdue University, West Lafayette, Indiana 47907, USA.
authors.forename: Diya
authors.initials: D
authors.lastname: Ren
authors.forename: Natalia A
authors.initials: NA
authors.lastname: Penner
authors.forename: Benjamin E
authors.initials: BE
authors.lastname: Slentz
authors.forename: Hamid
authors.initials: H
authors.lastname: Mirzaei
authors.forename: Fred
authors.initials: F
authors.lastname: Regnier
mesh_terms: D000818:Animals; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods; D003300:Copper; D006639:Histidine / Q000737:chemistry*; D006801:Humans; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D013387:Succinic Anhydrides
chemical_list: D010455:Peptides; D013387:Succinic Anhydrides; D006639:Histidine; C031801:succinic anhydride; D003300:Copper