Combining combinatorial chemistry and affinity chromatography: Highly selective inhibitors of human betaine - Homocysteine S-methyltransferase

Michaela Collinsová, Carmen Castro, Timothy A. Garrow, Athanasios Yiotakis, Vincent Dive, Jiří Jiráček

Research output: Contribution to journalArticlepeer-review

Abstract

A new method to find novel protein targets for ligands of interest is proposed. The principle of this approach is based on affinity chromatography and combinatorial chemistry. The proteins within a crude rat liver homogenate were allowed to interact with a combinatorial library of phosphinic pseudopeptides immobilized on affinity columns. Betaine: homocysteine S-methyltransferase (BHMT) was one of the proteins that was retained and subsequently eluted from these supports. The phosphinic pseudopeptides, which served as immobilized ligands for the isolation of rat BHMT, were then tested for their ability to inhibit human recombinant BHMT in solution. The most potent inhibitor also behaved as a selective ligand for the affinity purification of BHMT from a complex media. Further optimization uncovered Val-Phe-ψ[PO2--CH2]-Leu-His-NH 2 as a potent BHMT inhibitor that has an IC50 of about 1 μM.

Original languageEnglish (US)
Pages (from-to)113-122
Number of pages10
JournalChemistry and Biology
Volume10
Issue number2
DOIs
StatePublished - Feb 1 2003

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

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