Rotational diffusion of mitochondrial ADP/ATP carrier studied by saturation-transfer electron spin resonance.
The rotational mobility of the mitochondrial ADP/ATP carrier has been studied solubilized in Triton micelles, reincorporated in phospholipid liposomes, and in mitochondria. Spin-labeled analogues of the noncovalent inhibitors carboxyatractyloside and atractyloside were found to be strongly immobilized when bound to the carrier [Munding, A., Beyer, K., & Klingenberg, M. (1983) Biochemistry 22, 1941-1947], such that saturation-transfer electron spin resonance spectroscopy could be used to study the rotational motion of the protein. Spin-labeled maleimide covalently bound to the carrier was found to have independent segmental motion and hence to be unsuitable for studies of protein rotation. The ESR spectra of the carboxyatractyloside and atractyloside spin labels were found to contain a second component from label in the lipid or detergent, necessitating the use of the saturation-transfer ESR integral method [Horváth, L. I., & Marsh, D. (1983) J. Magn. Reson. 54, 363-373] to analyze the rotational motion of the label component bound to the protein. Effective rotational correlation times obtained from integration of the high-field region of the spectrum were lower than those obtained from the total spectral integral, indicating strongly anisotropic rotational diffusion of the carrier in the vesicular and membrane systems, with the spin-label z axis oriented preferentially perpendicular to the rotation axis.(ABSTRACT TRUNCATED AT 250 WORDS)
Horváth
L I
LI
Abteilung Spektroskopie, Max-Planck-Institut für Biophysikalische Chemie, Göttingen, Federal Republic of Germany.
Munding
A
A
Beyer
K
K
Klingenberg
M
M
Marsh
D
D
D000818:Animals; D001278:Atractyloside / Q000031:analogs & derivatives; D002417:Cattle; D004058:Diffusion; D004578:Electron Spin Resonance Spectroscopy; D008081:Liposomes; D008301:Maleimides; D008929:Mitochondria, Heart / Q000201:enzymology; D000226:Mitochondrial ADP, ATP Translocases; D009713:Nucleotidyltransferases; D013113:Spin Labels
D008081:Liposomes; D008301:Maleimides; D013113:Spin Labels; D001278:Atractyloside; C043592:maleimide; D000226:Mitochondrial ADP, ATP Translocases; D009713:Nucleotidyltransferases; C003853:carboxyatractyloside