Barnard M L, Gurdian S, Diep D, Ladd M, Turrens J F
Department of Biomedical Sciences, College of Allied Health Professions, University of South Alabama, Mobile 36688.
Arch Biochem Biophys. 1993 Feb 1;300(2):651-6. doi: 10.1006/abbi.1993.1090.
Incubation of 4% bovine serum albumin (BSA) with 1 mM tert-butyl hydroperoxide (t-BOOH) resulted in a peak of chemiluminescence followed by decay to a steady-state level of 18 counts per second above control. When using BSA of differing fatty acid content, the intensity of the initial peak was proportional to fatty acid content, while the steady-state chemiluminescence was independent of lipid content and depended only on BSA concentration. Light emission was inhibited by superoxide dismutase, desferrioxamine, and the antioxidant U-78518F. Oxidation of BSA by neutrophils activated with phorbol myristate acetate also increased chemiluminescence, in a process inhibitable by superoxide dismutase and U-78518F. When adding 1 mM t-BOOH in the presence of 20 microM heme to tryptophan, tyrosine, or to a lesser extent histidine, chemiluminescence correlated with increased oxygen consumption and the appearance of carbonyl derivatives, suggesting that chemiluminescence is a result of the decay to ground level of excited carbonyls. Lysine and glycine, on the other hand, were not oxidized to carbonyls after exposure to t-BOOH and did not emit light or consume significant oxygen during this challenge.
4%牛血清白蛋白（BSA）与1 mM叔丁基过氧化氢（t-BOOH）孵育会产生一个化学发光峰值，随后衰减至比对照高每秒18次计数的稳态水平。当使用不同脂肪酸含量的BSA时，初始峰值的强度与脂肪酸含量成正比，而稳态化学发光与脂质含量无关，仅取决于BSA浓度。超氧化物歧化酶、去铁胺和抗氧化剂U-78518F可抑制发光。用佛波酯肉豆蔻酸酯激活的中性粒细胞对BSA的氧化也会增加化学发光，此过程可被超氧化物歧化酶和U-78518F抑制。当在20 microM血红素存在的情况下向色氨酸、酪氨酸或在较小程度上向组氨酸中添加1 mM t-BOOH时，化学发光与耗氧量增加和羰基衍生物的出现相关，这表明化学发光是激发态羰基衰减至基态的结果。另一方面，赖氨酸和甘氨酸在暴露于t-BOOH后不会被氧化成羰基，在此过程中也不会发光或消耗大量氧气。
