Structural features of transiently modified beta-lactoglobulin relevant to the stable binding of large hydrophobic molecules

Evgenia Lozinsky, Stefania Iametti, Alberto Barbiroli, Gertz I. Likhtenshtein, Tamás Kálai, Kálmán Hideg, Francesco Bonomi

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Binding sites for hydrophobic molecules on bovine β-lactoglobulin, and their susceptibility to temperature, were studied by using various spectroscopic probes. Binding of probes carrying a single fluorophore moiety, a single nitroxide moiety, or both moieties on the same molecule, was followed by EPR and fluorescence. The presence of a fatty acid side chain in the dual probes was found to be required for binding to β-lactoglobulin. Binding occurred only after the protein was heated at temperatures below the threshold for its irreversible denaturation. Binding became extremely tight and stable upon cooling of the protein-probe mixture. Comparison among the various probes suggests that multiple binding sites for hydrophobes are present in the native protein, and in the partially-and reversibly-modified form of β-lactoglobulin present in solution at neutral pH and subdenaturing temperatures. Thus, the specificity of hydrophobes binding to β-lactoglobulin may be modulated by simple physical treatment of the protein.

Original languageEnglish
Pages (from-to)1-15
Number of pages15
JournalProtein Journal
Volume25
Issue number1
DOIs
Publication statusPublished - Jan 1 2006

Keywords

  • Dual fluorescence-spin probes
  • Hydrophobic binding sites
  • Temperature-induced protein modification
  • β-lactoglobulin

ASJC Scopus subject areas

  • Analytical Chemistry
  • Bioengineering
  • Biochemistry
  • Organic Chemistry

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