[HTML][HTML] The C331A mutant of neuronal nitric-oxide synthase is defective in arginine binding
P Martásek, RT Miller, Q Liu, LJ Roman… - Journal of Biological …, 1998 - ASBMB
It has been proposed that Cys 99 of human endothelial nitric oxide synthase (eNOS) is
responsible for tetrahydrobiopterin (BH 4) binding. To examine this possibility rigorously, we
expressed rat neuronal NOS (nNOS) in Escherichia coli, with the homologous Cys 331 to
Ala mutation, and characterized structural and functional attributes of the purified, mutated
enzyme. C331A-nNOS, as isolated, was catalytically incompetent. Upon prolonged
incubation with l-arginine (l-Arg), not only BH 4 binding but also catalytic activity could be …
responsible for tetrahydrobiopterin (BH 4) binding. To examine this possibility rigorously, we
expressed rat neuronal NOS (nNOS) in Escherichia coli, with the homologous Cys 331 to
Ala mutation, and characterized structural and functional attributes of the purified, mutated
enzyme. C331A-nNOS, as isolated, was catalytically incompetent. Upon prolonged
incubation with l-arginine (l-Arg), not only BH 4 binding but also catalytic activity could be …