Proteomic analysis of mouse kidney peroxisomes: identification of RP2p as a peroxisomal nudix hydrolase with acyl-CoA diphosphatase activity

Abstract

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.

Multiple Nudix family proteins possess mRNA decapping activity

“…1), demonstrating that decapping is not a general feature of 8-OH-dGTPases. Mouse Nudt19 (RP2p) was identified as a CoA diphosphatase (Ofman et al 2006). Similar to Nudt12, Nudt19 also contains a C-terminus type 1 peroxisomal targeting signal and may also localize to peroxisomes.…”

Biochemistry and genetics of inherited disorders of peroxisomal fatty acid metabolism

“…More detailed 17 Human ACOT4 combines the activities of three separate peroxisomal murine esterase encoded by Acot3 , Acot4 , and Acot5 (174). investigations showed that NUDT7 acts also acyl-CoAs, both medium-and long-chain acyl-CoAs, even choloyl-CoA and pristanoyl-CoA ( 191,192 ). The physiological role of NUDT7 is unclear.…”

Pex35 is a regulator of peroxisome abundance

“…In recent years this has been achieved by both genomewide screens and dedicated proteomic analyses. For example, proteomics screens of peroxisomes were able to identify new peroxisomal proteins in fungi (Marelli et al, 2004;Schäfer et al, 2001; Thoms et al, 2008;Yi et al, 2002), plants (Eubel et al, 2008;Fukao et al, 2002;Reumann et al, 2007Reumann et al, , 2009) and mammalian tissues, such as rat liver (Islinger et al, 2007(Islinger et al, , 2010; Kikuchi et al, 2004), mouse liver and kidney (Mi et al, 2007;Ofman et al, 2006;Wiese et al, 2007), and human liver (Gronemeyer et al, 2013). In addition, several genome-wide genetic screens in yeast have shed light on new proteins and their potential functions.…”