Binding of spin-labeled fatty acids and lysophospholipids to hydrophobic region of calmodulin.
In the presence of bovine brain calmodulin activated by calcium, the sharp triplet electron spin resonance (ESR) lines of free doxyl stearic acids decreased, and the broad resonance lines increased concomitantly, suggesting that the doxyl stearic acids bound to calmodulin calcium-dependently. The bound molecules were displaced by a calmodulin inhibitor, W-7, whereas their nitroxide radicals were hardly reduced by ascorbic acid, suggesting that the spin-labeled fatty acids bind to hydrophobic regions of calmodulin, and consequently inhibit calmodulin-dependent phosphodiesterase activity. These binding characteristics to calmodulin were different from those to bovine serum albumin. Moreover, the ESR spectra of two spin-labeled derivatives of lysophospholipid having a spin-labeled acyl group or a spin-labeled polar head group showed that it is the acyl chain of lysophospholipid that interacts with the hydrophobic region of calmodulin. The interactions of fatty acids and lysophospholipids with calmodulin seem to be quite different from those of acidic phospholipids, described previously [Suzuki, T., Katoh, H., & Uchida, M.K. (1986) Biochim. Biophys. Acta, 873, 379-386]. Thus, from the results of ESR study, we can obtain information on the function of fatty acids and lysophospholipids on calmodulin. Instead of enzyme assay, ESR spectroscopy is a useful means to examine lipid-protein interaction.
Suzuki-Nishimura
T
T
Department of Molecular Pharmacology, Meiji College of Pharmacy, Tokyo.
Uchida
M K
MK
D000818:Animals; D001921:Brain; D002147:Calmodulin / Q000737:chemistry / Q000378:metabolism*; D002417:Cattle; D003497:Cyclic N-Oxides; D004578:Electron Spin Resonance Spectroscopy; D008246:Lysophospholipids / Q000378:metabolism*; D011485:Protein Binding; D013113:Spin Labels / Q000138:chemical synthesis; D013229:Stearic Acids / Q000378:metabolism*
D002147:Calmodulin; D003497:Cyclic N-Oxides; D008246:Lysophospholipids; D013113:Spin Labels; D013229:Stearic Acids; C003959:TEMPO