Abstract
Publisher Summary This chapter discusses the experimental study, focusing on nicotinic acid adenine dinucleotide phosphate (NAADP + ). The nicotinic acid analog of NADP + is readily formed via the base-exchange reaction, catalyzed by beef spleen NAD(P) glycohydrolase. NAADP + formed is purified by chromatography on a column of Dowex-1-formate. The column is equilibrated, with 0.15 M formic acid, and the reaction mixture is injected in 1-ml aliquots, while the column is eluted, with 0.15 M formic acid, at a flow rate of 1 ml/min and the effluent diverted to drain. Under these conditions, nicotinic acid is flushed from the column, while NAADP + and other products accumulate. Multiple injections can be made, with little loss of sample, by keeping the reaction mixture in a calibrated centrifuge tube and drawing out only that quantity needed to fill the sample loop. The results show that NAADP + , the product of the base-exchange reaction, is present in greatest quantity (50.2%), followed by the product of the competing hydrolysis reaction, 2'P-ADPR (27.3%). The other products are relatively minor components. NAADP + is assayed by means of its conversion to NAAD + with alkaline phosphatase. The reaction product is then analyzed chromatographically.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
