Abstract
Natriuretic substances are a group of molecules affecting sodium homeostasis in the body. Recently, two new molecules having natriuresis effects, xanthurenic acid 8-O-β-d-glucoside and xanthurenic acid 8-O-sulfate (XA sulfate), have been isolated from human urine. In the present study, we have investigated the sulfation of xanthurenic acid (XA) in mouse tissues to assess the contribution of specific sulfotransferases (STs) to the reaction. Cytosols from tissues of both sexes of C57BL/6N mice (liver, stomach, jejunum, colon, and kidney) were capable of forming XA sulfate, with various Km values. Jejunum cytosol showed the lowest Km value, and its Vmax/Km value was much greater than those of other tissues. The kinetic analyses with recombinant mouse (m) STs (Sult1a1, Sult1b1, Sult1c2, and Sult1d1) showed the lowest Km value for mSult1b1, and the value was comparable with that for jejunum cytosol. The highest expression of mSult1b1 in small intestine was confirmed at the mRNA and protein levels. mSult1b1 is thus suggested as a major enzyme responsible for XA sulfation in jejunum. Similar to mSult1b1, human SULT1B1 and rat Sult1b1 mediated XA sulfation efficiently. Thus, XA is likely to be an endogenous substrate for ST1B members. In contrast to XA, an XA-related compound, kynurenic acid strongly inhibited mSult1b1-mediated sulfations, with IC50 values at a micromolar range. These results indicate the functional role of ST1B subfamily of ST in XA sulfate formation in the body.
Footnotes
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↵1 The nomenclature of human ST forms is based on the Human Genome Nomenclature Committee. For the rodent STs, the nomenclature system used in the National Center for Biotechnology Information is applied in this article, accompanying the names in the nomenclature system proposed by us with “ST” as a prefix (Nagata and Yamazoe, 2000; Nagata et al., 2005) for the first appearance, because rodent STs do not always have human counterparts. The accession numbers of STs are as follows: St1a4/mSult1a1, NM_133670; St1b3/mSult1b1, NM_019878; St1c9/mSult1c2, NM_026935; St1d1/mSult1d1, NM_016771; ST1A3/hSULT1A1, NM_001055; ST1A5/hSULT1A3, NM_003166; ST1B2/hSULT1B1, NM_014465; ST1C2/hSULT1C2, NM_001056; and ST1B1/rSult1b1, NM_022513.
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This work was supported in part by grant-in-aid from Ministry of Education, Culture, Sports, Sciences and Technology; Ministry of Health, Labor, and Welfare of Japan; and Comprehensive Research and Education Center for Planning of Drug Development and Clinical Education, Tohoku University 21st Century “Center of Excellence” Program.
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Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
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doi:10.1124/jpet.108.143164.
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ABBREVIATIONS: XA, xanthurenic acid; ST, sulfotransferase; PAPS, 3′-phosphoadenosine-5′-phosphosulfate; DTT, dithiothreitol; DOPAC, 3,4-dihydroxyphenylacetic acid; T3, l-triiodothyronine; BSA, bovine serum albumin; Ni-NTA, nickel-nitrilotriacetic acid; PAGE, polyacrylamide gel electrophoresis; m, mouse; PCR, polymerase chain reaction; r, rat; h, human; RT, reverse transcription.
- Received July 9, 2008.
- Accepted September 23, 2008.
- The American Society for Pharmacology and Experimental Therapeutics
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