Polyethenoadenosine phosphate as a fluorogenic substrate for barnase.
Polyethenoadenosine phosphate is rapidly hydrolyzed by barnase with an accompanying large increase in fluorescence. At pH 8, 0.2 M ammonium acetate, the initial rate of fluorescence increase is proportional to barnase concentration and thus provides an excellent quantitative assay for its activity. The preparation and use of this substrate in the barnase assay is presented in detail.
Fitzgerald
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PC
Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892.
Hartley
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RW
D001426:Bacterial Proteins; D000242:Cyclic AMP / Q000031:analogs & derivatives* / Q000737:chemistry; D005453:Fluorescence; D006868:Hydrolysis; D015394:Molecular Structure; D011061:Poly A; D011131:Polyribonucleotides / Q000737:chemistry*; D012260:Ribonucleases / Q000737:chemistry*; D013379:Substrate Specificity
D001426:Bacterial Proteins; D011131:Polyribonucleotides; D011061:Poly A; C005163:poly(1,N(6)-ethenoadenylic acid); D000242:Cyclic AMP; D012260:Ribonucleases; C019760:Bacillus amyloliquefaciens ribonuclease