the specific activity of recombinant soluble zf12-LOX mutant is increased by nearly 75% (25 nmol/min/mg) in the presence of calcium as compared to that in its absence (7 nmol/min/mg), while for the site-2 mutant, it is 5 nmol/min/mg irrespective of the presence of calcium. Ca2+ decreases the aggregation temperature of soluble zf12-LOX mutant, while secondary structure is maintained. Presence of Ca2+ induces increase in molecular dimensions of soluble zf12-LOX mutant
construction of a soluble version of zf12-LOX by mutagenesis. Mutation of the putative calcium-responsive amino acids in N-PLAT domain of soluble zf12-LOX and analysis of the oligomeric state, stability, structural integrity and conformational changes of zf12-LOX in response to calcium. Soluble zf12-LOX and the N-PLAT domain-mutant both proteins exist as compact monomers in solution, and the enzyme activity of soluble zf12-LOX is significantly increased in presence of calcium, the stimulatory effect of calcium on zf12-LOX is related to a change in protein structure. In contrast, enzyme with a mutated calcium regulatory site has reduced activity-response to calcium and restricted large re-modeling, suggesting that it retains a closedstate in response to calcium. Ca2+-dependent regulation is associated with different domain conformation(s) that might change the accessibility to substrate-binding site in response to calcium
construction of a soluble version of zf12-LOX by mutagenesis. Mutation of the putative calcium-responsive amino acids in N-PLAT domain of soluble zf12-LOX and analysis of the oligomeric state, stability, structural integrity and conformational changes of zf12-LOX in response to calcium. Soluble zf12-LOX and the N-PLAT domain-mutant both proteins exist as compact monomers in solution, and the enzyme activity of soluble zf12-LOX is significantly increased in presence of calcium, the stimulatory effect of calcium on zf12-LOX is related to a change in protein structure. In contrast, enzyme with a mutated calcium regulatory site has reduced activity-response to calcium and restricted large re-modeling, suggesting that it retains a closedstate in response to calcium. Ca2+-dependent regulation is associated with different domain conformation(s) that might change the accessibility to substrate-binding site in response to calcium