BUSCADOR RECOLECTA

Digital.CSIC. Repositorio Institucional del CSIC

oai:digital.csic.es:10261/310815

1 figure., (A) Coupled reaction used to measure ITPase activity. HAM1 ITPase hydrolyses ITP in the presence of Mg+2 to produce IMP and pyrophosphate (PPi). IMP is then oxidized by inosine monophosphate dehydrogenase (IMPDH) in the presence of NAD to produce XMP and NADH2, which is directly monitored in real time at 340 nm. The ITPase activity (U/ml) can be calculated as shown. (B) Cropped polyacrylamide gel stained with coomasie blue that illustrates the yield of MBP-HAM1 in elution fractions. The indicated MBP-HAM1 versions in fraction two, which had the higher protein concentration, are shown. (C) Kinetics of ITP hydrolysis, measured by the formation of NADH2 as indicated in (A), when 1 μl of a 1:10 dilution of the elution fractions shown in (B) were used in the reaction mix., Peer reviewed