Molecular cloning, expression and characterization of a novel mouse SULT6 cytosolic sulfotransferase.
By searching the mouse EST database, we identified a novel mouse cytosolic sulfotransferase (SULT) cDNA (RIKEN cDNA 2410078J06). Sequence analysis revealed that this new SULT belongs to the cytosolic SULT6 gene family. The recombinant form of this newly identified SULT, designated SULT6B1, was expressed using the pGEX-4T-1 glutathione S-transferase fusion system and purified from transformed BL21 Escherichia coli cells. Purified mouse SULT6B1 exhibited sulfonating activity toward thyroxine and bithionol among a variety of endogenous and xenobiotic compounds tested as substrates. pH optimum of purified mouse SULT6B1 was determined to be 8.0. Tissue-specific expression of mouse and human SULT6B1 was examined by RT-PCR. While human SULT6B1 was specifically expressed in kidney and testis, mouse SULT6B1 was detected in brain, heart, kidney, thymus, lung, liver and testis. Further studies are needed in order to clarify the role of SULT6B1 in the metabolism of thyroxine and possibly some xenobiotics in mouse.
Takahashi
Saki
S
Department of Biochemistry, University of Miyazaki, Miyazaki, Miyazaki 889-2192, Japan.
Sakakibara
Yoichi
Y
Mishiro
Emi
E
Kouriki
Haruna
H
Nobe
Rika
R
Kurogi
Katsuhisa
K
Yasuda
Shin
S
Liu
Ming-Cheh
MC
Suiko
Masahito
M
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001735:Bithionol / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D020224:Expressed Sequence Tags; D005260:Female; D015870:Gene Expression; D015723:Gene Library; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009206:Myocardium / Q000201:enzymology; D009928:Organ Specificity; D010724:Phosphoadenosine Phosphosulfate / Q000378:metabolism; D010802:Phylogeny; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D015238:Sulfotransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013974:Thyroxine / Q000378:metabolism
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D010724:Phosphoadenosine Phosphosulfate; D001735:Bithionol; C544761:SULT6B1 protein, human; C544760:SULT6B1 protein, mouse; D015238:Sulfotransferases; D013974:Thyroxine