Summary
-
1.
Incubation of high specific activity adenosine-5′-phosphosulfate (AP35S) with crude enzyme fractions or purified APS-sulfotransferase from wildtype and from a mutant which does not grow on sulfate (Sat2 -) leads to binding of label to protein as judged by separation on Sephadex G-25. Crude extracts of mutants Sat1 -, Sat6 -, and Sat7 -R1 lacking APS-sulfotransferase show 10 fold lower binding, indicating that APS-sulfotransferase activity is required.
-
2.
Protein-bound radioactivity is not released by mild acid and exchanges with sulfite excluding bound APS; this suggests a linkage of the organic thiosulfate type (R-S-SO3 -). Labeled protein releases radioactive SO3 2- and GSSO3 - with gluthathione (GS-); with BAL or S2-, SSO3 2- is released.
-
3.
The reaction of APS with APS-sulfotransferase is heat labile, but once bound, the radioactivity is still released by thiols from the heated protein.
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4.
Aged purified APS-sulfotransferase loses binding activity which is restored by adding the supernatant from heated fresh extract, indicating the participation of a cofactor, consistent with the view that a low molecular weight carrier participates in the main pathway of reduction.
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Abbreviations
- S2- :
-
sulfide
- SO3 2- :
-
sulfite
- G-SSO3 - :
-
glutathione S-sulfonate
- DTT:
-
dithiothreitol
- BAL:
-
2,3-dimercaptopropanol
- SSO3 2- :
-
thiosulfate
- APS:
-
adenosine-5′-phosphosulfate
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Abrams, W.R., Schiff, J.A. Studies of sulfate utilization by algae. Archiv. Mikrobiol. 94, 1–10 (1973). https://doi.org/10.1007/BF00414074
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DOI: https://doi.org/10.1007/BF00414074