Properties and regulation of adenosine 5'-phosphosulfate sulfotransferase from suspension cultures ofNicotiana sylvestris.

The properties and the regulation of adenosine 5'-phosphosulfate sulfotransferase extracted from cell suspension cultures ofNicotiana sylvestris was investigated. Optimal adenosine 5'-phosphosulfate sulfotransferase activity was obtained from the cells by extraction with 0.1 M tris-HCl, pH8.0, containing 2 M MgSO4 and 10 mM dithioerythritol. The K m for adenosine 5'-phosphosulfate in the sulfotransferase reaction was about 11 μM. Adenosine 5'-phosphosulfate in concentrations above 50 μM were inhibitory. The extratable adenosine 5'-phosphosulfate sulfotransferase activity decreased during cultivation with sulfate as the sole sulfur source, but after about 3 days it reached a constant level (50 to 100 nmol activated sulfate transferred h(-1) mg(-1) protein) which was maintained for at least 24 h. Addition of 0.5 mM cysteine to the culture medium decreased the extractable adenosine 5'-phosphosulfate sulfotransferase activity and blocked growth completely. With 0.1 mM cysteine an enzyme level of about 10% of the initial value was reached within 6 to 12 h without significant inhibition of growth. The added cysteine was absorbed rapidly and after 24 h cysteine could no longer be detected in the medium. Before the cysteine was completely depleted, the activity of adenosine 5'-phosphosulfate sulfotransferase started to increase, reaching ultimately a level which was comparable to the initial value.