Human platelet phenol sulfotransferase: partial purification and detection of two forms of the enzyme.

P. R. Butler, Robert J. Anderson, D. L. Venton

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Abstract

To begin to study the usefulness of platelet phenol sulfotransferase (PST) as a possible measure of the enzyme activity in other organs such as the brain, we purified human platelet PST 36-120-fold. Activity toward 3-methoxy-4-hydroxyphenylglycol (MHPG), dopamine, 5-hydroxytryptamine (5-HT), and phenol eluted in the same Sephadex G-100 and Affi-Gel Blue column fractions. Specific activities of the enzyme with MHPG, dopamine, 5-HT, and phenol as substrates were 1198, 1068, 401, and 408 units/mg protein, respectively. Optimal assay conditions were established for each substrate. Apparent Km values were 598 microM, 21 microM, 19 microM, and 500 microM for MHPG, dopamine, phenol, and 5-HT, respectively. Apparent Km values for 3'-phosphoadenosine-5'-phosphosulfate (PAPS) with the same four substrates ranged from 0.11 to 0.25 microM. The pH optima were 6.3 for phenol, 6.8 for dopamine, and 7.0 for MHPG and 5-HT. An additional pH optimum at 8.6 was present for 5-HT. A thermolabile form of the enzyme measured with dopamine and 5-HT, as well as a thermostable form measured with phenol, were present. Dichloronitrophenol (10(-5) M) noncompetitively inhibited the thermostable enzyme activity by 96% but decreased the thermolabile activity by only 36%. These studies provide the basis for a more accurate comparison of human platelet PST with the enzyme in the human brain and in other tissues.

Original languageEnglish
Pages (from-to)630-639
Number of pages10
JournalJournal of Neurochemistry
Volume41
Issue number3
StatePublished - Sep 1983
Externally publishedYes

Fingerprint

Arylsulfotransferase
Platelets
Purification
Phenol
Serotonin
Blood Platelets
Dopamine
Enzymes
Cibacron Blue F 3GA
Enzyme activity
Brain
Substrates
Phosphoadenosine Phosphosulfate
Assays
Tissue
4-hydroxyphenethylene glycol

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Human platelet phenol sulfotransferase : partial purification and detection of two forms of the enzyme. / Butler, P. R.; Anderson, Robert J.; Venton, D. L.

In: Journal of Neurochemistry, Vol. 41, No. 3, 09.1983, p. 630-639.

Research output: Contribution to journalArticle

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