A simple fluorometric method for cAMP: application to studies of brain adenylate cyclase activity.
A simple fluorometric method for the determination of cAMP is presented. The fluorescent derivative is 1,N6-etheno cyclic 3,5-monophosphate (etheno-cAMP). Maximal formation of this derivative occurs after reacting cAMP with chloroacetaldehyde for 15 minutes at 100 degrees C. Fluorescent derivatives are also produced from compounds which contain a 6-amino purine. The specificity of the method resides in the use of a reverse phase/HPLC system. The derivatization as well as the fluorescent response of etheno-cAMP is linear between 2.5 and 700 picomoles of cAMP. Studies of brain adenylate cyclase by the fluorometric/HPLC method indicated that this method is comparable to the established radioenzymatic method. Thus, the present method provides a simple rapid nonradioactive means for the determination of adenylate cyclase activity.
D000079:Acetaldehyde / Q000031:analogs & derivatives; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000201:enzymology*; D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid; D000242:Cyclic AMP / Q000031:analogs & derivatives* / Q000032:analysis* / Q000378:metabolism; D006168:Guinea Pigs; D013050:Spectrometry, Fluorescence
C001626:1,N(6)-ethenoadenosine 3',5'-monophosphate; C004656:chloroacetaldehyde; D000242:Cyclic AMP; D000262:Adenylyl Cyclases; D000079:Acetaldehyde