Evaluating immobilized metal affinity chromatography for the selection of histidine-containing peptides in comparative proteomics

Diya Ren, Natalia A. Penner, Benjamin E. Slentz, Hamid Mirzaei, Fred Regnier

Research output: Contribution to journalArticle

38 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)321-329
Number of pages9
JournalJournal of Proteome Research
Volume2
Issue number3
DOIs
StatePublished - May 2003

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Affinity chromatography
Affinity Chromatography
Histidine
Proteomics
Metals
Peptides
Amines
Aminoacylation
Acylation
Sepharose
Copper
Recovery

Keywords

  • Comparative proteomics
  • Cu(II)-IMAC
  • Peptide acylation

ASJC Scopus subject areas

  • Genetics
  • Biotechnology
  • Biochemistry

Cite this

Evaluating immobilized metal affinity chromatography for the selection of histidine-containing peptides in comparative proteomics. / Ren, Diya; Penner, Natalia A.; Slentz, Benjamin E.; Mirzaei, Hamid; Regnier, Fred.

In: Journal of Proteome Research, Vol. 2, No. 3, 05.2003, p. 321-329.

Research output: Contribution to journalArticle

Ren, Diya ; Penner, Natalia A. ; Slentz, Benjamin E. ; Mirzaei, Hamid ; Regnier, Fred. / Evaluating immobilized metal affinity chromatography for the selection of histidine-containing peptides in comparative proteomics. In: Journal of Proteome Research. 2003 ; Vol. 2, No. 3. pp. 321-329.
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