[Measurement of NOS activity by monitoring the conversion of 3H-arginine].
OBJECTIVE
To establish a method for measuring the constitutive and inducible nitric oxide synthase (NOS) through the conversion of L-3H arginine to L-3H citrulline.


METHODS
NOS extracted from cerebellar of porcine and rat was purificated through DEAE cellulose or/and 2', 5', -ADP agarose. NOS activity was assayed by monitoring the conversion of 3H-arginine to 3H-citrulline.


RESULTS
A 4,460 fold purification of the porcine cerebellar NOS was obtained with an activity of 669 pmol mg-1 min-1 and a 2.7% recovery. For purification of rat cerebellar NOS, affinity chromatography with 2', 5'-ADP agarose column provided a 2646 fold purification of enzyme activity with a 688 pmol.ml-1.min-1 and 27% recovery. The purified rat and porcine cerebellar NOS constituted a single band on SDS/PAGE at about 160,000.


CONCLUSIONS
A stable and special method for measuring NOS activity was established.
Qiang
W
W
Institute of Basic Medical Science, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing.
Liu
J
J
Lu
F
F
D000818:Animals; D001120:Arginine / Q000032:analysis; D002846:Chromatography, Affinity; D002956:Citrulline / Q000032:analysis; D005260:Female; D008297:Male; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
D002956:Citrulline; D001120:Arginine; D019001:Nitric Oxide Synthase