Purified nitric oxide synthase from rat brain
AssigneeUniversity of Iowa Research Foundation
InventorsJohn P. N. Rosazza, Yijun Chen
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Abstract
A nitric oxide synthase (NOS) protein is obtained from rat brain and purified to a specific activity at least 6,360-fold that of a crude tissue preparation. The NOS protein has a denatured MW of 105 kD determined by SDS-PAGE, a dimeric MW of 230 kD determined by gel filtration and requires FAD, FMN, Ca.sup.2+ and tetrahydropterin for the production of nitric oxide from L-arginine or an arginine-rich peptide, oligopeptide or protein substrate.
Claims7
A nitric oxide synthase (NOS) protein (nNOS-II), purified from rat brain, said protein having a denatured molecular mass as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing conditions of about 105 kD, and a native homodimeric molecular mass as determined by gel filtration of about 230 kD, requiring FAD, FMN, Ca.sup.2+ and tetrahydrobiopterin cofactors for the production of nitric oxide either from L-arginine, or an analog or derivative thereof, or from an arginine-rich peptide, oligopeptide, or protein substrate, and further providing that the NOS protein is calmodulin-dependent with L-arginine as a substrate and calmodulin-independent with an arginine-rich polypeptide as a substrate,
- wherein the NOS protein is purified to specific activity at least 6,360-fold that of a crude tissue preparation.
Dependencies are inferred from the claim text.