U-3'-BCIP: a chromogenic substrate for the detection of RNase A in recombinant DNA expression systems.
The synthesis of the bovine pancreatic ribonuclease A (RNase A, EC 3.1.27.5) chromogenic substrate uridine-3'-(5-bromo-4-chloroindol-3-yl)-phosphate (U-3'-BCIP) is described. RNase A catalyzes the hydrolysis of U-3'-BCIP to release a halogenated indol-3-ol that undergoes rapid aerobic oxidation to the dark blue 5,5'-dibromo-4,4'-dichloroindigo. Preliminary kinetic studies indicate that this compound may have practical use for assaying RNase A activity both in vitro and in vivo, e.g. in screening bacterial colonies for RNase A produced by recombinant DNA methods.
Witmer
M R
MR
Baker Laboratory of Chemistry, Cornell University, Ithaca, NY 14853-1301.
Falcomer
C M
CM
Weiner
M P
MP
Kay
M S
MS
Begley
T P
TP
Ganem
B
B
Scheraga
H A
HA
D000818:Animals; D002417:Cattle; D004274:DNA, Recombinant; D007700:Kinetics; D015394:Molecular Structure; D010084:Oxidation-Reduction; D012259:Ribonuclease, Pancreatic / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D014542:Uridine Monophosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism
D004274:DNA, Recombinant; C068167:uridine-3'-(5-bromo-4-chloroindol-3-yl)-phosphate; D014542:Uridine Monophosphate; D012259:Ribonuclease, Pancreatic