Nitric oxide synthase (NOS) was isolated from the uterus of adult female rats by diethylaminoethyl (DEAE) column and further purified by 2',5'-ADP agarose. The chromatographic properties revealed two isoenzymes, NOS1 and NOS2. The molecular weights of both isoenzymes was approximately 155 Kd by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE) which was similar to NOS1 and NOS2 from rat brain. The enzymes required nicotinamide adenine dinucleotide phosphate (NADPH), Ca+2 and calmodulin as cofactors. However, in the absence of calmodulin and/or calcium NOS1 was reduced by approximately 96%, while NOS2 was reduced by approximately 70%. This maximal enzyme activity was similar for brain. These results demonstrate that two isoforms of NOS are present in the rat uterus.
Bibliographical noteFunding Information:
This work was supported in part Vikings Children6 Fund.
- Female reproduction
- Nitric oxide synthase
- Sprague-Dawley rat