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, T. Kalai, K. Hideg, Francesco Bonomi

Research output: Contribution to journalArticle

18 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 2006

Fingerprint

Lactoglobulins
Molecules
Proteins
Temperature
Binding sites
Binding Sites
Denaturation
Fluorophores
Fatty Acids
Fatty acids
Fluorescence
Paramagnetic resonance
Cooling

Keywords

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

ASJC Scopus subject areas

  • Biochemistry

Cite this

Structural features of transiently modified beta-lactoglobulin relevant to the stable binding of large hydrophobic molecules. / Lozinsky, Evgenia; Iametti, Stefania; Barbiroli, Alberto; Likhtenshtein, Gertz I.; Kalai, T.; Hideg, K.; Bonomi, Francesco.

In: Protein Journal, Vol. 25, No. 1, 01.2006, p. 1-15.

Research output: Contribution to journalArticle

Lozinsky, Evgenia ; Iametti, Stefania ; Barbiroli, Alberto ; Likhtenshtein, Gertz I. ; Kalai, T. ; Hideg, K. ; Bonomi, Francesco. / Structural features of transiently modified beta-lactoglobulin relevant to the stable binding of large hydrophobic molecules. In: Protein Journal. 2006 ; Vol. 25, No. 1. pp. 1-15.
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