The polypeptide Syn67 interacts physically with human holocarboxylase synthetase, but is not a target for biotinylation

Yousef I. Hassan, Hideaki Moriyama, Janos Zempleni

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Holocarboxylase synthetase (HCS) catalyzes the binding of biotin to lysines in carboxylases and histones in two steps. First, HCS catalyzes the synthesis of biotinyl-5′-AMP; second, the biotinyl moiety is ligated to lysine residues. It has been proposed that step two is fairly promiscuous, and that protein biotinylation may occur in the absence of HCS as long as sufficient exogenous biotinyl-5′-AMP is provided. Here, we identified a novel polypeptide (Syn67) with a basic patch of lysines and arginines. Yeast-two-hybrid assays and limited proteolysis assays revealed that both N- and C-termini of HCS interact with Syn67. A potential target lysine in Syn67 was biotinylated by HCS only after arginine-to-glycine substitutions in Syn67 produced a histone-like peptide. We identified a Syn67 docking site near the active pocket of HCS by in silico modeling and site-directed mutagenesis. Biotinylation of proteins by HCS is more specific than previously assumed.

Original languageEnglish (US)
Pages (from-to)35-41
Number of pages7
JournalArchives of Biochemistry and Biophysics
Volume495
Issue number1
DOIs
StatePublished - Mar 2010

Keywords

  • Biotin
  • Domains
  • Holocarboxylase synthetase
  • Substrate
  • Syn67

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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