Bovine serum albumin. Study of the fatty acid and steroid binding sites using spin labeled lipids

J. D. Morrisett, H. J. Pownall, A. M. Gotto

Research output: Contribution to journalArticle

88 Scopus citations

Abstract

Three spin labeled derivatives of stearic acid and two derivatives of palmitic acid have been used to study the structure of the strong fatty acid binding site of bovine serum albumin. The steroid and indole binding sites have been studied using spin labeled derivatives of androstol and indole, respectively. Paramagnetic resonance and fluorescence quenching data suggest that the fatty acid, steroid and indole binding sites may be identical. The mobility of the nitroxyl group at C 8 of palmitic acid bound to albumin at a 1:1 molar ratio is unaffected when the carboxyl group is esterified. When the nitroxyl group is located at C 5 on this acid, its motion is detectably increased by esterification of the carboxyl group but the magnitude of this change is small. This result suggests that the carboxyl group may play a minor role in the binding of fatty acids to the strongest fatty acid binding site of albumin. When stearic acid derivatives bearing the nitroxide at C 5, C 12 and C 16 are bound to albumin at a ligand to albumin ratio of 1, the order of mobility at 0 to 30° is C 16 > C 12 C 5. Although motion at the methyl terminus is always greater than at the COOH terminus in the range 0 to 60°, a simple monotonic increase in chain motion between the two termini is not observed. Arrhenius plots of the motion parameters for these bound fatty acids show two abrupt changes in slope. The temperature ranges for these changes are 15 to 23° and 38 to 45°. These results suggest that when one mole of spin labeled fatty acid is bound to albumin, the protein undergoes a conformational change in each of these temperature ranges.

Original languageEnglish (US)
Pages (from-to)2487-2494
Number of pages8
JournalJournal of Biological Chemistry
Volume250
Issue number7
StatePublished - 1975

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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