Isolation and characterisation of a novel rabbit sulfotransferase isoform belonging to the SULT1A subfamily.
Sulfotransferases (SULTs) catalyse the sulfonation of both endogenous and exogenous compounds including hormones, catecholamines, drugs and xenobiotics. While in most occasions, sulfonation is a detoxication pathway, in the case of certain drugs and carcinogens, it leads to metabolic activation. Since, the rabbit has been extensively used for both pharmacological and toxicological studies, the purpose of this study was to further characterise the sulfotransferase system of this animal. In the present study, a novel sulfotransferase isoform (GenBank Accession no. AF360872) was isolated from a rabbit liver cDNA lambdaZAP II library. The full-length sequence of the clone was 1138 bp long and contained a coding region of 888 bp encoding a cytosolic protein of 295 amino acids (deduced molecular weight 34,193 Da). The amino acid sequence of this novel SULT isoform showed >70% identity with members of the SULT1A subfamily of sulfotransferases from other species. Upon expression of the encoded rabbit sulfotransferase in Escherchia coli (E. coli), it was shown that the enzyme was capable of sulfonating both p-nitrophenol (K(m) and Vmax values of 0.15 microM and 897.5 nmol/min/mg protein, respectively) and dopamine (K(m) and V(max) values of 175.3 microM and 151.1 nmol/min/mg protein, respectively). Based on the sequence data obtained and substrate specificity, this new rabbit sulfotransferase was named rabSULT1A1. Immunoblotting was used to demonstrate that rabSULT1A1 protein is expressed in liver, duodenum, jejunum, ileum, colon and rectum.
Riley
E
E
Department of Physiology and Pharmacology, School of Biomedical Sciences, University of Queensland, QLD 4072, Brisbane, Australia
Bolton-Grob
R
R
Liyou
N
N
Wong
C
C
Tresillian
M
M
McManus
M E
ME
D000595:Amino Acid Sequence; D000818:Animals; D015239:Arylsulfotransferase; D001483:Base Sequence; D015153:Blotting, Western / Q000379:methods; D003001:Cloning, Molecular; D018076:DNA, Complementary; D006801:Humans; D007527:Isoenzymes / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D015238:Sulfotransferases / Q000145:classification / Q000235:genetics / Q000378:metabolism*
D018076:DNA, Complementary; D007527:Isoenzymes; D015238:Sulfotransferases; D015239:Arylsulfotransferase; C117544:SULT1A1 protein, human