Kurogi Katsuhisa, Shimohira Takehiko, Kouriki-Nagatomo Haruna, Zhang Guisheng, Miller Ethan R, Sakakibara Yoichi, Suiko Masahito, Liu Ming-Cheh
Department of Pharmacology, College of Pharmacy and Pharmaceutical Sciences, The University of Toledo, 3000 Arlington Avenue, Toledo, OH 43614, USA.
Department of Biochemistry and Applied Biosciences, University of Miyazaki, 1-1, Gakuenkibanadai-Nishi Miyazaki 889-2192, Japan.
J Biochem. 2017 Dec 1;162(6):403-414. doi: 10.1093/jb/mvx044.
The cytosolic sulphotransferase SULT1C3 remained the most poorly understood human SULT. The SULT1C3 gene has been shown to contain alternative exons 7 and 8, raising the question concerning their evolutionary origin and implying the generation of multiple SULT1C3 variants. Two SULT1C3 splice variants, SULT1C3a and SULT1C3d, were investigated to verify the impact of alternative C-terminal sequences on their sulphating activity. Sequence homology and gene location analyses were performed to verify the orthology of the SULT1C3 gene. The SULT1C3 gene appears to be present only in humans and other primates, but alternative exons 7b and 8b share high degrees of homology with corresponding regions of rodent SULT1C1 genes, implying their evolutionary origin being from a defunct human SULT1C1 gene. Purified recombinant SULT1C3a and SULT1C3d were analyzed for sulphating activities toward a variety of endogenous and xenobiotic compounds. While SULT1C3a displayed weaker activities and strict substrate specificity toward hydroxyl-chlorinated biphenyls, SULT1C3d exhibited broader substrate specificity toward bile acids and thyroid hormones as well as hydroxyl-chlorinated biphenyls. Molecular docking simulation suggested that Tyr249 and Met257 may play an important role in substrate recognition by SULT1C3d. Alternative splicing of exons 7 and 8 sequences resulted in differential catalytic properties of SULT1C3 variants.
胞质磺基转移酶SULT1C3仍然是人们了解最少的人类磺基转移酶。已表明SULT1C3基因包含选择性外显子7和8,这就引发了关于它们进化起源的问题,并意味着会产生多种SULT1C3变体。对两种SULT1C3剪接变体SULT1C3a和SULT1C3d进行了研究,以验证C端序列的差异对其硫酸化活性的影响。进行了序列同源性和基因定位分析,以验证SULT1C3基因的直系同源性。SULT1C3基因似乎仅存在于人类和其他灵长类动物中,但选择性外显子7b和8b与啮齿动物SULT1C1基因的相应区域具有高度同源性,这意味着它们的进化起源是一个已失效的人类SULT1C1基因。分析了纯化的重组SULT1C3a和SULT1C3d对多种内源性和外源性化合物的硫酸化活性。虽然SULT1C3a对羟基氯化联苯的活性较弱且底物特异性严格,但SULT1C3d对胆汁酸、甲状腺激素以及羟基氯化联苯表现出更广泛的底物特异性。分子对接模拟表明,Tyr249和Met257可能在SULT1C3d识别底物中起重要作用。外显子7和8序列的选择性剪接导致了SULT1C3变体的催化特性不同。