Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.14.15.33 extracted from

  • Li, S.; Ouellet, H.; Sherman, D.H.; Podust, L.M.
    Analysis of transient and catalytic desosamine-binding pockets in cytochrome P-450 PikC from Streptomyces venezuelae (2009), J. Biol. Chem., 284, 5723-5730.
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

Crystallization (Comment) Organism
structures of the PikC D50N mutant cocrystallized with narbomycin, to 1.85 A resolution, and with 10-deoxymethymycin, to 3.2 A resolution. In the mutant D50N, the desosamine moiety of both 10-deoxymethymycin and narbomycin is bound in a catalytically productive buried site, suggesting a two-step substrate binding mechanism, whereby desosamine is recognized in the two subsites to allow the macrolide substrate to sequentially progress toward a catalytically favorable orientation Streptomyces venezuelae

Protein Variants

Protein Variants Comment Organism
D50N increase in catalytic activity. Binding affinity for 10-desoxymethymycin increases about 4fold, binding affinity for narbomycin about 1.3fold compared to wild-type. 60% conversion of 10-desoxymethymycin, ratio of products neomethymycin to methymycin is 1.15. 44.4% conversion of substrate narbomycin Streptomyces venezuelae
D50N/E85Q 18.4% conversion of 10-desoxymethymycin, ratio of products neomethymycin to methymycin is 0.38. 3.5% conversion of substrate narbomycin Streptomyces venezuelae
D50N/E94Q no conversion of 10-desoxymethymycin. 5.3% conversion of substrate narbomycin Streptomyces venezuelae
E85Q 18% conversion of 10-desoxymethymycin, ratio of products neomethymycin to methymycin is 0.26. 8.1% conversion of substrate narbomycin Streptomyces venezuelae
E94Q no conversion of 10-desoxymethymycin. 5.4% conversion of substrate narbomycin Streptomyces venezuelae

Organism

Organism UniProt Comment Textmining
Streptomyces venezuelae O87605
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
10-deoxymethymycin + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 i.e. YC-17. 40% conversion, ratio of products neomethymycin to methymycin is 0.81 Streptomyces venezuelae methymycin + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
10-deoxymethymycin + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 40% conversion, ratio of products neomethymycin to methymycin is 0.81 Streptomyces venezuelae neomethymycin + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
narbomycin + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 binding kinetics are biphasic and accurately described by a double exponential function revealing a fast first phase followed by a slow second one. The kinetic rate of the first binding step, accounting for about 50% of the reaction amplitude, is fast and protein-dependent. The rate of the second step is slow, about 13 per s Streptomyces venezuelae pikromycin + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O 35% conversion ?