Bun-ya M, Maebuchi M, Kamiryo T, Kurosawa T, Sato M, Tohma M, Jiang L L, Hashimoto T
Faculty of Integrated Arts and Sciences, Hiroshima University, Higashi-Hiroshima.
J Biochem. 1998 Feb;123(2):347-52. doi: 10.1093/oxfordjournals.jbchem.a021943.
The formation of cholic acid and chenodeoxycholic acid through cleavage of the side chains of CoA esters of 3alpha,7alpha,12alpha-trihydroxy-5beta-choles tan-26-oic acid and 3alpha,7alpha-dihydroxy-5beta-cholestan-26-oic acid is believed to occur in peroxisomes. Recently, we found a new peroxisomal enzyme, D-3-hydroxyacyl-CoA dehydratase/D-3-hydroxyacyl-CoA dehydrogenase bifunctional protein, and suggested that this bifunctional protein is responsible for the conversion of 3alpha,7alpha,12alpha-trihydroxy-5beta-cholest-2 4-en-26-oyl-CoA and 3alpha,7alpha-dihydroxy-5beta-cholest-24-en-26-oyl-CoA to their 24-oxo-forms. In the present study, the products of this bifunctional protein reaction were analyzed by gas chromatography-mass spectrometry, and the formation of 24-oxo-27-nor-cholestanes was confirmed. Previously, we found a new thiolase in Caenorhabditis elegans, P-44, and suggested that P-44 and sterol carrier protein x, a peroxisomal protein, constitute a second group of 3-oxoacyl-CoA thiolases. The production of cholic acid and chenodeoxycholic acid from the precursors on incubation with the bifunctional protein and sterol carrier protein x or P-44 was confirmed by gas chromatography.
3α,7α,12α-三羟基-5β-胆甾烷-26-酸和3α,7α-二羟基-5β-胆甾烷-26-酸的辅酶A酯侧链裂解形成胆酸和鹅去氧胆酸,这一过程被认为发生在过氧化物酶体中。最近,我们发现了一种新的过氧化物酶体酶,D-3-羟酰基辅酶A脱水酶/D-3-羟酰基辅酶A脱氢酶双功能蛋白,并提出这种双功能蛋白负责将3α,7α,12α-三羟基-5β-胆甾-24-烯-26-酰基辅酶A和3α,7α-二羟基-5β-胆甾-24-烯-26-酰基辅酶A转化为它们的24-氧代形式。在本研究中,通过气相色谱-质谱联用分析了这种双功能蛋白反应的产物,并证实了24-氧代-27-降胆甾烷的形成。此前,我们在秀丽隐杆线虫中发现了一种新的硫解酶P-44,并提出P-44和过氧化物酶体蛋白甾醇载体蛋白x构成了第二组3-氧代酰基辅酶A硫解酶。通过气相色谱法证实了在与双功能蛋白和甾醇载体蛋白x或P-44一起孵育时,前体物质可产生胆酸和鹅去氧胆酸。