Ethan Miller1, Munaf H Zalzala1,2, Maryam S Abunnaja1, Katsuhisa Kurogi1,3, Yoichi Sakakibara3, Masahito Suiko3, Ming-Cheh Liu4. 1. Department of Pharmacology, College of Pharmacy and Pharmaceutical Sciences, University of Toledo Health Science Campus, 3000 Arlington Avenue, Toledo, OH, 43614, USA. 2. Department of Pharmacology and Toxicology, College of Pharmacy, University of Baghdad, Baghdad, Iraq. 3. Department of Biochemistry and Applied Biosciences, University of Miyazaki, Miyazaki, 889-2192, Japan. 4. Department of Pharmacology, College of Pharmacy and Pharmaceutical Sciences, University of Toledo Health Science Campus, 3000 Arlington Avenue, Toledo, OH, 43614, USA. ming.liu@utoledo.edu.
Abstract
BACKGROUND AND OBJECTIVES: Previous studies have demonstrated the metabolism of tibolone through sulfation, with the cytosolic sulfotransferase (SULT) SULT2A1 as the major responsible enzyme. The current study aimed to investigate how SULT2A1 genetic polymorphisms may affect the dehydroepiandrosterone (DHEA)- and tibolone-sulfating activity of SULT2A1. METHODS: Site-directed mutagenesis was employed to generate cDNAs encoding ten different SULT2A1 allozymes. Recombinant SULT2A1 allozymes were expressed in BL21 E. coli cells, and purified using glutathione-sepharose affinity chromatography. An established sulfotransferase assay was used to analyze DHEA- and tibolone-sulfating activity of the purified SULT2A1 allozymes. RESULTS: The nine human SULT2A1 allozymes plus the wild-type SULT2A1 were found to display differential sulfating activity toward DHEA and tibolone. Kinetic analysis revealed that different SULT2A1 allozymes exhibited differential substrate affinity and catalytic efficiency toward the two substrates tested. CONCLUSION: The results obtained provided useful information concerning the differential metabolism of tibolone through sulfation in individuals with different SULT2A1 genotypes.
BACKGROUND AND OBJECTIVES: Previous studies have demonstrated the metabolism of tibolone through sulfation, with the cytosolic sulfotransferase (SULT) SULT2A1 as the major responsible enzyme. The current study aimed to investigate how SULT2A1 genetic polymorphisms may affect the dehydroepiandrosterone (DHEA)- and tibolone-sulfating activity of SULT2A1. METHODS: Site-directed mutagenesis was employed to generate cDNAs encoding ten different SULT2A1 allozymes. Recombinant SULT2A1 allozymes were expressed in BL21 E. coli cells, and purified using glutathione-sepharose affinity chromatography. An established sulfotransferase assay was used to analyze DHEA- and tibolone-sulfating activity of the purified SULT2A1 allozymes. RESULTS: The nine humanSULT2A1 allozymes plus the wild-type SULT2A1 were found to display differential sulfating activity toward DHEA and tibolone. Kinetic analysis revealed that different SULT2A1 allozymes exhibited differential substrate affinity and catalytic efficiency toward the two substrates tested. CONCLUSION: The results obtained provided useful information concerning the differential metabolism of tibolone through sulfation in individuals with different SULT2A1 genotypes.
Authors: K Yanagisawa; Y Sakakibara; M Suiko; Y Takami; T Nakayama; H Nakajima; K Takayanagi; Y Natori; M C Liu Journal: Biosci Biotechnol Biochem Date: 1998-05 Impact factor: 2.043
Authors: Eva Lundström; Alexander Christow; Wendy Kersemaekers; Gunilla Svane; Edward Azavedo; Gunnar Söderqvist; Mirjam Mol-Arts; Jan Barkfeldt; Bo von Schoultz Journal: Am J Obstet Gynecol Date: 2002-04 Impact factor: 8.661
Authors: Katsuhisa Kurogi; Mohammed I Rasool; Fatemah A Alherz; Amal A El Daibani; Ahsan F Bairam; Maryam S Abunnaja; Shin Yasuda; Lauren J Wilson; Ying Hui; Ming-Cheh Liu Journal: Expert Opin Drug Metab Toxicol Date: 2021-06-30 Impact factor: 4.936