ELECTRON-PARAMAGNETIC-RESONANCE INVESTIGATIONS OF FREE RADICAL-INDUCED ALTERATIONS IN NEOCORTICAL SYNAPTOSOMAL MEMBRANE-PROTEIN INFRASTRUCTURE

被引:87
作者
HENSLEY, K
CARNEY, J
HALL, N
SHAW, W
BUTTERFIELD, DA
机构
[1] UNIV KENTUCKY, DEPT CHEM, LEXINGTON, KY 40506 USA
[2] UNIV KENTUCKY, CTR MEMBRANE SCI, LEXINGTON, KY 40506 USA
[3] UNIV KENTUCKY, DEPT PHARMACOL, LEXINGTON, KY 40536 USA
关键词
SYNAPTOSOMES; FREE RADICALS; EPR; SPIN LABELING; AGING; ALZHEIMERS DISEASE;
D O I
10.1016/0891-5849(94)90018-3
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
Evidence is presented that free radical stress can directly induce physico-chemical alterations in rodent neocortical synaptosomal membrane proteins. Synaptosomes were prepared from gerbil cortical brain tissue and incubated with 3 mM ascorbate and various concentrations of exogenous Fe2+ for 30-240 min at 37 degrees C. Synaptosomes were then lysed and covalently labeled with the protein thiol-selective spin label MAL-6 (2,2,6,6-tetramethyl-4-maleimidopiperdin-1-oxyl) and subjected to electron paramagnetic resonance (EPR) spectrometry. In separate experiments. synaptosomal membranes were labeled with the thiol-specific spin label MTS ((1-oxyl-2,2,5,5-tetramethyl-pyrroline-3-methyl)-methanethiosulfonate), or the lipid-specific spin probe 5-NS (5-nitroxide stearate). Free radical stress induced by iron/ascorbate treatment has a rigidizing effect on the protein infrastructure of these membranes, as appraised by EPR analysis of membrane protein-bound spin label, bat no change was detected in the lipid component of the membrane. These results are discussed with reference to potential oxidative mechanisms in aging and neurological disorders.
引用
收藏
页码:321 / 331
页数:11
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