Ethanesulfonic acid, 2-(((3alpha,5beta,7alpha,12alpha)-3,7,12-trihydroxy-24-oxocholan-24-yl)amino)-

• 分子结构分类: 有机化合物 - 脂质和类脂质分子 - 类固醇和类固醇衍生物
• 英文名称: Ethanesulfonic acid, 2-(((3alpha,5beta,7alpha,12alpha)-3,7,12-trihydroxy-24-oxocholan-24-yl)amino)-
• 英文别名: 2-[[(4R)-4-[(3R,5S,7R,8R,9S,10S,12S,13R,14S,17R)-3,7,12-trihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]pentanoyl]amino]ethanesulfonate
• 化学式: C26H44NO7S-
• 分子量: 514.7
• InChiKey: WBWWGRHZICKQGZ-HZAMXZRMSA-M

计算性质

• 辛醇/水分配系数(LogP)：
  2.1
• 重原子数：
  35
• 可旋转键数：
  6
• 环数：
  4.0
• sp3杂化的碳原子比例：
  0.96
• 拓扑面积：
  155
• 氢给体数：
  4
• 氢受体数：
  7

反应信息

• 作为反应物：
  描述：

  PAPS 、 Ethanesulfonic acid, 2-(((3alpha,5beta,7alpha,12alpha)-3,7,12-trihydroxy-24-oxocholan-24-yl)amino)- 生成 Adenosine 3',5'-bismonophosphate(4-) 、 氢(+1)阳离子 、 taurocholate 7-sulfate
  参考文献：
  名称：

  His48在小鼠胞质磺基转移酶SULT2A8中对胆汁酸的7α-羟基硫酸化的关键作用。

  摘要：

  已知胞质磺基转移酶（SULT）SULT2A亚家族的成员严重参与类固醇和胆汁酸的稳态。SULT2A8是7α-羟基胆汁酸偏爱的小鼠SULT，已被确定为负责小鼠特异性7-O-胆汁酸硫酸化的主要酶。有趣的是，SULT2A8在第99位缺少保守的催化His残基。因此，尚不清楚SULT2A8介导的胆汁酸7-O-硫酸化的催化机理。在这项研究中，我们进行了突变分析，以便深入了解这个尚未解决的问题。获得的结果表明，两个氨基酸残基His48和Leu99对小鼠SULT2A8而言是唯一的，但对其他SULTs而言则不是，对于其对胆汁酸的7-O硫酸化活性至关重要。

  DOI：

  10.1080/09168451.2018.1464897
• 作为产物：
  描述：

  Cholate 、 牛磺酸 生成 水 、 Ethanesulfonic acid, 2-(((3alpha,5beta,7alpha,12alpha)-3,7,12-trihydroxy-24-oxocholan-24-yl)amino)-
  参考文献：
  名称：

  长双歧杆菌胆汁盐水解酶的生化和遗传学表征。

  摘要：

  从长双歧杆菌SBT2928分离出胆汁盐水解酶（BSH），纯化并鉴定。此外，我们首次描述了双歧杆菌属成员中编码BSH（bsh）的基因的克隆和分析。根据推导的氨基酸序列测定，该酶的天然分子量为125,000-130,000，亚单位分子量为35,024，表明该酶是四聚体。长双歧杆菌BSH的最适pH为5至7，最适温度为40℃。该酶被硫醇酶抑制剂强烈抑制，表明Cys残基可能参与催化反应。长双歧杆菌的BSH可以水解所有六种主要的人胆汁盐和至少两种动物胆汁盐。基于特异性和K（m）值，检测到对甘氨酸缀合的胆汁酸略有偏爱。确定了bsh的核苷酸序列，并将其用于同源性研究，转录本分析以及各种突变体的构建和分析。与其他细菌的BSH和球形芽孢杆菌的青霉素V酰基转移酶（PVA）的同源性很高。基于已经阐明晶体结构的BSH和PVA的相似性，可以将BSH分类为N末端亲核水解酶，而将Cys作为N末端氨基酸。通过定点诱变进行的

  DOI：

  10.1128/aem.66.6.2502-2512.2000

文献信息

• The Human Bile Acid-CoA:Amino Acid N-Acyltransferase Functions in the Conjugation of Fatty Acids to Glycine
  作者：James O'Byrne、Mary C. Hunt、Dilip K. Rai、Masayumi Saeki、Stefan E.H. Alexson

  DOI：10.1074/jbc.m300987200

  日期：2003.9

  Bile acid-CoA:amino acid N-acyltransferase (BACAT) catalyzes the conjugation of bile acids to glycine and taurine for excretion into bile. By use of site-directed mutagenesis and sequence comparisons, we have identified Cys-235, Asp-328, and His-362 as constituting a catalytic triad in human BACAT (hBACAT) and identifying BACAT as a member of the type I acyl-CoA thioesterase gene family. We therefore

  胆汁酸-CoA：氨基酸N-酰基转移酶（BACAT）催化胆汁酸与甘氨酸和牛磺酸的结合，从而排泄到胆汁中。通过使用定点诱变和序列比较，我们已鉴定出Cys-235，Asp-328和His-362构成人BACAT（hBACAT）中的催化三联体，并将BACAT鉴定为I型酰基辅酶A硫酯酶基因家族。因此，我们假设hBACAT也可能将脂肪酰基辅酶A和/或共轭脂肪酸水解为甘氨酸。我们在这里显示重组hBACAT也可以水解长链和非常长链的饱和酰基CoAs（主要是C16：0-C26：0），并且通过质谱法验证了hBACAT还可以将脂肪酸缀合到甘氨酸上。组织表达研究表明，BACAT在肝，胆囊以及近端和远端肠中都有强表达。然而，BACAT还可以在与胆汁酸形成和运输无关的多种组织中表达，这表明在调节长链脂肪酸的细胞内水平方面也起着重要的作用。在人皮肤成纤维细胞中的绿色荧光蛋白定位实验表明，hBACAT酶主要是胞质的。因此
• Cloning and characterization of a conjugated bile acid hydrolase gene from Clostridium perfringens
  作者：J P Coleman、L L Hudson

  DOI：10.1128/aem.61.7.2514-2520.1995

  日期：1995.7

  The gene encoding a conjugated bile acid hydrolase (CBAH) from Clostridium perfringens 13 has been cloned and expressed in Escherichia coli, and its nucleotide sequence has been determined. Nucleotide and predicted amino acid sequence analyses indicated that the gene product is related to two previously characterized amidases, a CBAH from Lactobacillus plantarum (40% identity) and a penicillin V amidase from Bacillus sphaericus (34% identity). The product is apparently unrelated to a CBAH from C. perfringens for which N-terminal sequence information was determined. The gene product was purified from recombinant E. coli and used to raise antibody in rabbits. The presence of the protein in C. perfringens was then confirmed by immunoblot analysis. The protein was shown to have a native molecular weight of 147,000 and a subunit molecular weight of 36,100, indicating its probable existence as a tetramer. Disruption of the chromosomal C. perfringens CBAH gene with a chloramphenicol resistance cartridge resulted in a mutant strain which retained partial CBAH activity. Polyacrylamide gel electrophoresis followed by enzymatic activity staining and immunoblotting indicated that the mutant strain no longer expressed the cloned CBAH (CBAH-1) but did express at least one additional CBAH (CBAH-2). CBAH-2 was immunologically distinct from CBAH-1, and its mobility on native polyacrylamide gels was different from that of CBAH-1. Furthermore, comparisons of pH optima and substrate specificities of CBAH activities from recombinant E. coli and wild-type and mutant C. perfringens provided further evidence for the presence of multiple CBAH activities in C. perfringens.

  来自Clostridium perfringens 13的编码共轭胆汁酸水解酶（CBAH）基因已在大肠杆菌中克隆和表达，并确定了其核苷酸序列。核苷酸和预测的氨基酸序列分析表明，该基因产物与两种先前表征的酰胺酶有关，即来自Lactobacillus plantarum的CBAH（40％同源性）和来自Bacillus sphaericus的青霉素V酰胺酶（34％同源性）。该产物显然与确定N末端序列信息的C. perfringens的CBAH不相关。从重组E. coli中纯化了基因产物，并用于兔子中提高抗体。然后通过免疫印迹分析确认了C. perfringens中的蛋白质存在。该蛋白质显示具有147,000的天然分子量和36,100的亚基分子量，表明其可能存在为四聚体。使用氯霉素耐药性卡带破坏染色体C. perfringens CBAH基因导致突变株保留部分CBAH活性。聚丙烯酰胺凝胶电泳随后进行酶活性染色和免疫印迹分析表明，突变株不再表达克隆的CBAH（CBAH-1），但至少表达另外一种CBAH（CBAH-2）。CBAH-2在免疫学上与CBAH-1不同，并且其在天然聚丙烯酰胺凝胶上的迁移率与CBAH-1不同。此外，从重组E. coli和野生型和突变C. perfringens中的CBAH活性的pH最适值和底物特异性的比较，为C. perfringens中存在多种CBAH活性提供了进一步的证据。
• Cloning and sequencing of the 7 alpha-hydroxysteroid dehydrogenase gene from Escherichia coli HB101 and characterization of the expressed enzyme
  作者：T Yoshimoto、H Higashi、A Kanatani、X S Lin、H Nagai、H Oyama、K Kurazono、D Tsuru

  DOI：10.1128/jb.173.7.2173-2179.1991

  日期：1991.4

  The 7 alpha-hydroxysteroid dehydrogenase (EC 1.1.1.159) gene from Escherichia coli HB101 was cloned and expressed in E. coli DH1. The hybrid plasmid pSD1, with a 2.8-kbp insert of chromosomal DNA at the BamHI site of pBR322, was subcloned into pUC19 to construct plasmid pSD3. The entire nucleotide sequence of an inserted PstI-BamHI fragment of plasmid pSD3 was determined by the dideoxy chain-termination method. Within this sequence, the mature enzyme protein-encoding sequence was found to start at a GTG initiation codon and to comprise 765 bp, as judged by comparison with the protein sequence. The deduced amino acid sequence of the enzyme indicated that the molecular weight is 26,778. The transformant of E. coli DH1 harboring pSD3 with a 1.8-kbp fragment showed about 200-fold-higher enzyme activity than the host. The enzyme was purified by a single chromatography step on DEAE-Toyopearl and obtained as crystals, with an activity yield of 39%. The purified enzyme was homogeneous, as judged by sodium dodecyl sulfate gel electrophoresis. The enzyme was most active at pH 8.5 and stable between pH 8 and 9. The enzyme was NAD+ dependent and had a pI of 4.3. The molecular mass was estimated to be 120 kDa by the gel filtration method and 28 kDa by electrophoresis, indicating that the enzyme exists in a tetrameric form.

  大肠杆菌HB101的7α-羟类固醇脱氢酶（EC 1.1.1.159）基因被克隆并在大肠杆菌DH1中表达。将含有2.8-kbp染色体DNA插入到pBR322的BamHI位点的杂交质粒pSD1亚克隆到pUC19中构建出质粒pSD3。通过dideoxy链终止法确定了质粒pSD3的插入PstI-BamHI片段的整个核苷酸序列。在该序列中，成熟酶蛋白编码序列从GTG启动密码子开始，由765个碱基组成，与蛋白质序列比较得出。酶的推导氨基酸序列表明分子量为26,778。携带1.8-kbp片段的大肠杆菌DH1转化体，其酶活性比宿主高约200倍。酶经DEAE-Toyopearl单一色谱步骤纯化，并以晶体形式获得，活性收率为39％。纯化的酶通过十二烷基硫酸钠凝胶电泳判断为均一。该酶在pH 8.5时最活跃，在pH 8和9之间稳定。该酶依赖于NAD+，其等电点为4.3。通过凝胶过滤法估计分子量为120 kDa，通过电泳法估计为28 kDa，表明该酶以四聚体形式存在。
• Falany C.N.; Johnson M.R.; Barnes S., J Biol Chem, 1994, 0021-9258, 19375-9
  作者：Falany C.N.、Johnson M.R.、Barnes S.、Diasio R.B.

  DOI：——

  日期：——
• Identification and characterization of a novel PPARα-regulated and 7α-hydroxyl bile acid-preferring cytosolic sulfotransferase mL-STL (Sult2a8)
  作者：Lu Feng、Yee-Lok Yuen、Jian Xu、Xing Liu、Martin Yan-Chun Chan、Kai Wang、Wing-Ping Fong、Wing-Tai Cheung、Susanna Sau-Tuen Lee

  DOI：10.1194/jlr.m074302

  日期：2017.6

  PPAR alpha has been known to play a pivotal role in orchestrating lipid, glucose, and amino acid metabolism via transcriptional regulation of its target gene expression during energy deprivation. Recent evidence has also suggested that PPAR alpha is involved in bile acid metabolism, but how PPAR alpha modulates the homeostasis of bile acids during fasting is still not clear. In a mechanistic study aiming to dissect the spectrum of PPAR alpha target genes involved in metabolic response to fasting, we identified a novel mouse gene (herein named mL-STL for mouse liver-sulfotransferase-like) that shared extensive homology with the Sult2a subfamily of a superfamily of cytosolic sulfotransferases, implying its potential function in sulfonation. The mL-STL gene expressed predominantly in liver in fed state, but PPAR alpha was required to sustain its expression during fasting, suggesting a critical role of PPAR alpha in regulating the mL-STL-mediated sulfonation during fasting. Functional studies using recombinant His-tagged mL-STL protein revealed its narrow sulfonating activities toward 7 alpha-hydroxyl primary bile acids, including cholic acid, chenodeoxycholic acid, and alpha-muricholic acid, and thus suggesting that mL-STL may be the major hepatic bile acid sulfonating enzyme in mice. Together, these studies identified a novel PPAR alpha-dependent gene and uncovered a new role of PPAR alpha as being an essential regulator in bile acid biotransformation via sulfonation during fasting.