Ca2+-induced increased lipid packing and domain formation in submitochondrial particles.: A possible early step in the mechanism of Ca2+-stimulated generation of reactive oxygen species by the respiratory chain

被引:150
作者
Grijalba, MT
Vercesi, AE
Schreier, S
机构
[1] Univ Sao Paulo, Inst Chem, Dept Biochem, BR-05599970 Sao Paulo, Brazil
[2] Univ Estadual Campinas, Sch Med, Dept Clin Pathol, BR-13083970 Campinas, SP, Brazil
关键词
D O I
10.1021/bi9828674
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Ca2+ and P-i accumulation by mitochondria triggers a number of alterations leading to nonspecific increase in inner membrane permeability [Kowaltowski, A. J., et al. (1996) J. Biol. Chem. 271, 2929-2934]. The molecular nature of the membrane perturbation that precedes oxidative damage is still unknown. EPR spectra of spin probes incorporated in submitochondrial particles (SMP) and in model membranes suggest that Ca2+-cardiolipin (CL) complexation plays an important role. Ca2+-induced lipid domain formation was detected in SMP but not in mitoplasts, in SMP extracted lipids, or in CL-containing liposomes. The results were interpreted in terms of Ca2+ sequestration of CL tightly bound to membrane proteins, in particular the ADP-ATP carrier, and formation of CE-enriched strongly immobilized clusters in lipid shells next to boundary lipid. The in-plane lipid and protein rearrangement is suggested to cause increased reactive oxygen species production in succinate-supplemented, antimycin A-poisoned SMP, favoring the formation of carbon-centered radicals, detected by EPR spin trapping. Removal of tightly bound CL is also proposed to cause protein aggregation, facilitating intermolecular thiol oxidation. Lipid peroxidation was also monitored by the disappearance of the nitroxide EPR spectrum. The decay was faster for nitroxides in a more hydrophobic environment, and was inhibited by butylated hydroxytoluene, by EGTA, or by substituting Mg2+ for Ca2+. In addition, Ca2+ caused an increase in permeability, evidenced by the release of carboxyfluorescein from respiring SMP. The results strongly support Ca2+ binding to CL as one of the early steps in the molecular mechanism of Ca2+-induced nonspecific inner mitochondrial membrane permeabilization.
引用
收藏
页码:13279 / 13287
页数:9
相关论文
共 54 条
[11]  
EDIDIN M, 1997, CURR OPIN STRUC BIOL, P582
[12]  
FAGIAN MM, 1990, J BIOL CHEM, V265, P19955
[13]  
FOLCH J, 1957, J BIOL CHEM, V226, P497
[14]   SPIN LABEL STUDY OF LOCAL ANESTHETIC-LIPID MEMBRANE INTERACTIONS - PHASE-SEPARATION OF THE UNCHARGED FORM AND BILAYER MICELLIZATION BY THE CHARGED FORM OF TETRACAINE [J].
FREZZATTI, WA ;
TOSELLI, WR ;
SCHREIER, S .
BIOCHIMICA ET BIOPHYSICA ACTA, 1986, 860 (03) :531-538
[15]   HYDROXYL RADICAL GENERATION DURING MITOCHONDRIAL ELECTRON-TRANSFER AND THE FORMATION OF 8-HYDROXYDESOXYGUANOSINE IN MITOCHONDRIAL-DNA [J].
GIULIVI, C ;
BOVERIS, A ;
CADENAS, E .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1995, 316 (02) :909-916
[16]   CALCIUM AND FREE-RADICALS IN HYPOXIA/REOXYGENATION INJURY OF RENAL EPITHELIAL-CELLS [J].
GREENE, EL ;
PALLER, MS .
AMERICAN JOURNAL OF PHYSIOLOGY, 1994, 266 (01) :F13-F20
[17]   Inhibition of membrane lipid peroxidation by a radical scavenging mechanism: a novel function for hydroxyl-containing ionophores [J].
Grijalba, MT ;
Andrade, PB ;
Meinicke, AR ;
Castilho, RF ;
Vercesi, AE ;
Schreier, S .
FREE RADICAL RESEARCH, 1998, 28 (03) :301-318
[18]   Oxidative stress, thiol reagents, and membrane potential modulate the mitochondrial permeability transition by affecting nucleotide binding to the adenine nucleotide translocase [J].
Halestrap, AP ;
Woodfield, KY ;
Connern, CP .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (06) :3346-3354
[19]   ASYMMETRICAL ORIENTATION OF PHOSPHOLIPIDS AND THEIR INTERACTIONS WITH MARKER ENZYMES IN PIG-HEART MITOCHONDRIAL INNER MEMBRANE [J].
HARB, JS ;
COMTE, J ;
GAUTHERON, DC .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1981, 208 (01) :305-318
[20]   VISUALIZATION OF CA-2+-INDUCED PHOSPHOLIPID DOMAINS [J].
HAVERSTICK, DM ;
GLASER, M .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1987, 84 (13) :4475-4479