Molecular Cloning, Expression, and Characterization of a Novel Mouse Liver SULT1B1 Sulfotransferase.
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概要
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A mouse liver homogenate was shown to contain enzymatic activities catalyzing the sulfation of 3, 4-dihydroxyphenylalanine (Dopa) and tyrosine isomers with a pH optimum of 8.25. Western blot analysis revealed a 34 kDa protein exhibiting immunologic crossreactivity to antiserum against rat liver SULT1B1 sulfotransferase. By employing the reverse transcriptase-polymerase chain reaction (RT-PCR) technique, a 910-base pair product encoding the putative mouse liver SULT1B1 sulfotransferase was obtained. Using this PCR product as a probe, a cDNA containing the entire open reading frame of the mouse liver SULT1B1 sulfotransferase was cloned from a mouse liver Lambda ZAP cDNA library. The nucleotide sequence indicated it is a new enzyme. The deduced amino acid sequence exhibited 87.6, 72.3, 55.9, 54.2, 52.8, 51.1, and 49.4% identity to the amino acid sequences of the rat liver SULT1B1 sulfotransferase, human thyroid hormone sulfotransferase, mouse phenol sulfotransferase, rat liver phenol sulfotransferase, rat liver hydroxyarylamine sulfotransferase, mouse estrogen sulfotransferase, and rat estrogen sulfotransferase. Upon transfection of COS-7 cells with an expression vector (pcDNA3) harboring the cDNA encoding this new enzyme, a 34 kDa protein exhibiting immunologic cross-reactivity to antiserum against the rat liver SULT1B1 sulfotransferase was expressed. The recombinant sulfotransferase exhibited enzymatic activities toward Dopa and tyrosine isomers, as well as dopamine and 3, 3', 5-triiodo-L-thyronine. Northern blot analyses indicated the SULT1B1 sulfotransferase was predominantly expressed in liver, but not in the other ten mouse organs examined. Furthermore, the enzyme was found to be expressed in a developmental stage-dependent manner, being at a very low level in liver samples from 1-day-old mice and then gradually increasing to the maximum level in liver samples from 4-week-old mice.
- 社団法人 日本生化学会の論文
著者
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Sakakibara Yoichi
Department Of Biochemistry And Applied Biosciences Faculty Of Agriculture University Of Miyazaki
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Saeki Yuichi
Department Of Agricultural Chemistry Faculty Of Agriculture Kyushu University
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Araki Yoichi
Department Of Biochemistry The University Of Texas Health Center
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Yanagisawa Ken
Department Of Biochemistry Ii Jichi Medical School
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NAKAJIMA Hiroshi
Department of Allergy and Clinical Immunology, Chiba University Hospital
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LIU Ming-Cheh
Department of Biochemistry, The University of Texas Health Center at Tyler
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SUIKO Masahito
Department of Agricultural Chemistry, Miyazaki University
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Sakakibara Yoichi
Department of Biochemistry, The University of Texas Health Center
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