Van Veldhoven P P, Vanhove G, Assselberghs S, Eyssen H J, Mannaerts G P
Afdeling Farmacologie, Katholieke Universiteit Leuven, Belgium.
J Biol Chem. 1992 Oct 5;267(28):20065-74.
Rat liver peroxisomes contain three acyl-CoA oxidases:palmitoyl-CoA oxidase, pristanoyl-CoA oxidase, and trihydroxycoprostanoyl-CoA oxidase. The three oxidases were separated by anion-exchange chromatography of a partially purified oxidase preparation, and the column eluate was analyzed for oxidase activity with different acyl-CoAs. Short chain mono (hexanoyl-) and dicarboxylyl (glutaryl-)-CoAs and prostaglandin E2-CoA were oxidized exclusively by palmitoyl-CoA oxidase. Long chain mono (palmitoyl-) and dicarboxylyl (hexadecanedioyl-)-CoAs were oxidized by palmitoyl-CoA oxidase and pristanoyl-CoA oxidase, the former enzyme catalyzing approximately 70% of the total eluate activity. The very long chain lignoceroyl-CoA was also oxidized by palmitoyl-CoA oxidase and pristanoyl-CoA oxidase, the latter enzyme catalyzing approximately 65% of the total eluate activity. Long chain 2-methyl branched acyl-CoAs (2-methylpalmitoyl-CoA and pristanoyl-CoA) were oxidized for approximately 90% by pristanoyl-CoA oxidase, the remaining activity being catalyzed by trihydroxycoprostanoyl-CoA oxidase. The short chain 2-methylhexanoyl-CoA was oxidized by trihydroxycoprostanoyl-CoA oxidase and pristanoyl-CoA oxidase (approximately 60 and 40%, respectively, of the total eluate activity). Trihydroxycoprostanoyl-CoA was oxidized exclusively by trihydroxycoprostanoyl-CoA oxidase. No oxidase activity was found with isovaleryl-CoA and isobutyryl-CoA. Substrate dependences of palmitoyl-CoA oxidase and pristanoyl-CoA oxidase were very similar when assayed with the same (common) substrate. Since the two oxidases were purified to a similar extent and with a similar yield, the contribution of each enzyme to substrate oxidation in the column eluate probably reflects its contribution in the intact liver.
大鼠肝脏过氧化物酶体含有三种酰基辅酶A氧化酶：棕榈酰辅酶A氧化酶、降植烷酰辅酶A氧化酶和三羟基胆甾烷酰辅酶A氧化酶。通过对部分纯化的氧化酶制剂进行阴离子交换色谱法分离这三种氧化酶，并用不同的酰基辅酶A分析柱洗脱液的氧化酶活性。短链单（己酰-）和二羧基（戊二酰-）辅酶A以及前列腺素E2-辅酶A仅被棕榈酰辅酶A氧化酶氧化。长链单（棕榈酰-）和二羧基（十六烷二酰-）辅酶A被棕榈酰辅酶A氧化酶和降植烷酰辅酶A氧化酶氧化，前一种酶催化约70%的总洗脱液活性。极长链木蜡酰辅酶A也被棕榈酰辅酶A氧化酶和降植烷酰辅酶A氧化酶氧化，后一种酶催化约65%的总洗脱液活性。长链2-甲基支链酰基辅酶A（2-甲基棕榈酰辅酶A和降植烷酰辅酶A）约90%被降植烷酰辅酶A氧化酶氧化，其余活性由三羟基胆甾烷酰辅酶A氧化酶催化。短链2-甲基己酰辅酶A被三羟基胆甾烷酰辅酶A氧化酶和降植烷酰辅酶A氧化酶氧化（分别约占总洗脱液活性的60%和40%）。三羟基胆甾烷酰辅酶A仅被三羟基胆甾烷酰辅酶A氧化酶氧化。异戊酰辅酶A和异丁酰辅酶A未发现氧化酶活性。用相同（共同）底物测定时，棕榈酰辅酶A氧化酶和降植烷酰辅酶A氧化酶的底物依赖性非常相似。由于这两种氧化酶的纯化程度和产率相似，每种酶对柱洗脱液中底物氧化的贡献可能反映了其在完整肝脏中的贡献。
