Spectrophotometric assay of NADase-catalyzed reactions |
| |
| Authors: | D A Yost B M Anderson |
| |
| Affiliation: | Department of Biochemistry and Nutrition, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061 USA |
| |
| Abstract: | A spectrophotometric method for the assay of NADase-catalyzed reactions was developed. The assay consisted of monitoring the decrease in absorbance at 275 nm accompanying the enzyme-catalyzed hydrolysis of ?-NAD. A millimolar extinction coefficient of 0.89 at 275 nm was determined for the hydrolysis of the nicotinamide-ribosidic bond of ?-NAD. Under assay conditions the assay was shown to be linear up to 50% completion. A linear relationship between the rate of ?-NAD hydrolyzed and the amount of NADase added was observed. The Km and Vmax values for Bungarus fasciatus venom NADase-catalyzed hydrolysis of ?-NAD were determined spectrophotometrically and were shown to be the same as those determined by other analytical methods. |
| |
| Keywords: | To whom correspondence should be addressed. |
| 本文献已被 ScienceDirect 等数据库收录！ |
