Abstract
Arylsulfotransferase catalyzes the transfer of a sulfate group from 3′-phosphoadenosine-5′-phosphosulfate (PAPS) to a phenolic acceptor substrate. We discovered a novel type of sulfotransferase from an anaerobic bacterium of human intestine, Eubacterium A-44. In the bacterial enzyme PAPS did not serve as a donor and all alcohols did not as acceptors. The new arylsulfotransferase was purified 185-fold from a crude extract of sonicated bacteria to homogeneity. The enzyme (MW 315 kd) was composed of four identical subunits (MW 80 kd) whose N-terminal amino acid was arginine, and its optimal pH and pI were 8–9 and 3.9, respectively. The enzyme catalyzed stoichiometric transfer of a sulfate group from a phenol sulfate ester to other phenols, with strict specificity. With tyramine as an acceptor, p-acetylphenyl sulfate was the best donor, followed by 4-methylumbelliferyl sulfate and p-nitrophenyl sulfate. With p-nitrophenyl sulfate as a donor, naphthol was the best acceptor, followed by estradiol, phenol, tyrosine methylester, tyramine, and epinephrine in decreasing order. Only the 4-position of catecholamines was specifically sulfated. Naturally occurring phenolic compounds, such as flavone, chalcone, and xanthone, were sulfated as well. Tyrosine-containing peptides were enzymatically sulfated: enkephalin, LH-RH, vasopressin, angiotensins, proctorin, CCK-8, and phyllocaerulein were sulfated with high yields. The novel sulfotransferase is expected to be applicable to enzymatic O-sulfation of tyrosine-containing hormones. The 35S-labeled sulfate group from (35S)p-nitrophenyl sulfate was incorporated into a tyrosyl residue at the active site of the enzyme (2 mole 35S/mole of enzyme). The enzyme was inactivated by diethylpyrocarbamate and TLCK, chemical modifying agents for a histidyl residue. The reaction mechanism of arylsulfotransferase was proposed as follows: a donor substrate combines a histidyl residue with concomitant release of a phenolic compound. The sulfate group of the histidyl residue transfers to a tyrosyl residue, and then to an acceptor with the binding of another donor substrate to the histidyl residue.
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References
Banerjee, R. K., and Roy, A. B. (1966). Mol. Pharmacol. 2, 56–66.
Jakoby, W. B., Sekura, R. D., Lyon, E. S., Marcus, C. J., and Wang, J. L. (1980). In Enzymatic Basis of Detoxication (Jakoby, W. B. ed.), Academic Press, New York, Vol. 2, pp. 199–228.
Kim, D.-H., Konishi, L., and Kobashi, K. (1986). Biochim. Biophys. Acta 872, 33–41.
Kobashi, K., Akao, T., Takebe, S., Fukaya, Y., and Kim, D.-H. (1984). Sulfur Amino Acids 7, 437–441.
Kobashi, K., Fukaya, Y., Kim, D.-H., Akao, T., and Takebe, S. (1986). Arch. Biochem. Biophys. 245, 537–539.
Roy, A. B. (1981). In Sulfation of Drugs and Related Compounds (Mulder, G. J. ed.), CRC Press, Boca Raton, Florida, pp. 83–130.
Sekura, R. D., Duffel, M. W., and Jakoby, W. B. (1981). Meth. Enzymol. 77, 197–206.
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This article was presented during the proceedings of the International Conference on Macromolecular Structure and Function, held at the National Defence Medical College, Tokorozawa, Japan, December 1985.
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Kobashi, K., Kim, DH. & Morikawa, T. A novel type of arylsulfotransferase. J Protein Chem 6, 237–244 (1987). https://doi.org/10.1007/BF00250287
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DOI: https://doi.org/10.1007/BF00250287