Proteomic analysis of mouse kidney peroxisomes: identification of RP2p as a peroxisomal nudix hydrolase with acyl-CoA diphosphatase activity
Proteomic analysis of mouse kidney peroxisomes: identification of RP2p as a peroxisomal nudix hydrolase with acyl-CoA diphosphatase activity
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DOI：
10.1042/bj20050893
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发表时间：
2006-01-15
影响因子：
4.1
通讯作者：
Wanders, RJA
中科院分区：
文献类型：
--
作者：
Ofman, R;Speijer, D;Wanders, RJA
通过对小鼠肾脏过氧化物体的蛋白质组学分析，鉴定出一种新的Nudex水解酶，命名为RP2p，由D7RP2e基因编码。RP2p由357个氨基酸组成，包含两个保守结构域：Nudex水解区和CoA结合区。此外，在C末端还发现了PTS(Peroxisomal靶向信号)1型(Ala-His-Leu)。酶性质分析表明，RP2p是一种辅酶A二磷酸酶，对辅酶A、氧化辅酶A和多种辅酶A酯具有活性，包括氯代辅酶A酯和支链脂肪酰辅酶A酯。RP2p的酶学性质表明，在低底物浓度下，中长链脂肪酰辅酶A酯是主要底物。在pH为9或以上时酶活力最高，需要有镁或锰离子存在。亚细胞分级研究表明，小鼠肾脏中的所有辅酶A二磷酸酶活性仅限于过氧化物体。
Proteomic analysis of mouse kidney peroxisomes resulted in the identification of a novel nudix hydrolase designated RP2p, which is encoded by the D7RP2e gene. RP2p consists of 357 amino acids and contains two conserved domains: a nudix hydrolase domain and a CoA-binding domain. In addition, a PTS (peroxisomal targeting signal) type 1 (Ala-His-Leu) was found at the C-terminus. Analysis of the enzyme characteristics revealed that RP2p is a CoA diphosphatase with activity towards CoA, oxidized CoA and a wide range of CoA esters, including choloyl-CoA and branched-chain fatty-acyl-CoA esters. The enzymatic properties of RP2p indicate that at low substrate concentrations medium and long-chain fatty-acyl-CoA esters are the primary substrates. Enzyme activity was optimal at pH 9 or above, and required the presence of Mg2+ or Mn2+ ions. Subcellular fractionation studies revealed that all CoA diphosphatase activity in mouse kidney is restricted to peroxisomes.