The response to ribulose bisphosphate(4-) (RuBP (4-)) and RuBP-Mg (2-) in catalysis by structurally divergent RuBP carboxylase/oxygenases.
Free ribulose bisphosphate (RuBP(4-)) rather than its magnesium complex (RuBP-Mg(2-)) was the apparent substrate for spinach ribulose bisphosphate carboxylase/oxygenase. The apparent Km for total RuBP (pH 8.0 at 30° C) increased with increasing Mg(2+) concentrations from 11.6 μM at 13.33 mM Mg(2+) to 32.6 μM at 40.33 mM Mg(2+). Similarly the apparent Km for RuBP-Mg(2-) complex increased with increasing Mg(2+) from 9.4 μM at 13.33 mM Mg(2+) to 29.7 μM at 40.33 mM Mg(2+). However, the Km values for uncomplexed RuBP(4-) were independent of the (saturating) concentration of Mg(2+) (Km=2.2 μM). The Vmax did not vary with the changing concentrations of Mg(2+).In contrast, the Km for total RuBP remained constant with varying Mg(2+) concentrations (Km=59.5 μM) for the enzyme from R. rubrum. The apparent Km for the RuBP-Mg(2-) complex decreased with increasing Mg(2+) concentrations from 16.0 μM at 7.5 mM Mg(2+) to 5.9 μM at 27.5 mM Mg(2+). The initial velocity for the C. vinosum enzyme was also found to be independent of the (saturating) concentration of Mg(2+) when total RuBP was varied in the assay. Thus the response to total RuBP by these two bacterial enzymes, which markedly differ in structure, was closely similar.
Biochemistry/Biophysics Program, Washington State University, 99164, Pullman, WA.
D J
DJ
Roach
B A
BA
McFadden