A protein binding assay for adenosine 3':5'-cyclic monophosphate.
نویسنده
چکیده
A simple and sensitive assay for adenosine 3':5'-cyclic monophosphate (cAMP) has been developed that is based on competition for protein binding of the nucleotide, presumably to a cAMP-dependent protein kinase. The nucleotide-protein complex is adsorbed on a cellulose ester filter. Assay conditions are such that a binding constant approaching 10(-9) M is obtained, and the assay is thus sensitive to 0.05-0.10 pmol of cAMP.
منابع مشابه
Attempts to detect cyclic adenosine 3':5'-monophosphate in higher plants by three assay methods.
Endogenous levels of cyclic adenosine-3':5'-monophosphate in coleoptile first leaf segments of oat (Avena sativa L.), potato (Solanum tuberosum L.) tubers, tobacco (Nicotiana tabacum L.) callus, and germinating seeds of lettuce (Lactuca sativa L.) were measured with a modified Gilman binding assay and a protein kinase activation assay. The incorporation of adenosine-8-(14)C into compounds with ...
متن کاملRadioimmunoassay for Cyclic Nucleotides
Sensitive and specific radioimmunoassays have been developed for cyclic adenosine 3’, 5’-monophosphate (CAMP), cyclic guanosine 3’ ,5’-monophosphate (cGMP), cyclic inosine 3’ ,5’-monophosphate (cIMP), and cyclic uridine 3’, 5’monophosphate (cUMP). These assays are based upon competition of the cyclic nucleotide with isotopically labeled cyclic nucleotide derivatives for binding sites on specifi...
متن کاملDifferential binding of cyclic adenosine 3' ,5'-monophosphate to the cyclic adenosine 3' ,5'-monophosphate receptor protein in Escherichia coli.
Binding of cyclic adenosine 3' ,5'-monophosphate (cAMP) by the cAMP receptor protein in crude cell-free extracts of Escherichia coli was characterized. When cell were grown in glucose, binding was inhibited 50% relative to extracts from cells grown with succinate as carbon source . This inhibition could be relieved by dialysis.
متن کاملFailure to Detect Cyclic 3', 5'-Adenosine Monophosphate in Healthy and Crown Gall Tumorous Tissues of Vicia faba.
Attempts were made to provide proof for the occurrence of cyclic 3',5'-adenosine monophosphate in healthy and crown gall tissues of Vicia faba. Although our purified extracts gave positive readings in the Gilman binding assay for cyclic AMP, they were not digested by a specific cyclic 3',5'-adenosine monophosphate phosphodiesterase from beef heart. The extracts were digested, however, by a part...
متن کاملIdentification and characterization of the adenosine 3',5'-cyclic monophosphate binding proteins appearing during the development of Dictyostelium discoideum.
A photosensitive, radioactive analogue of cyclic adenosine monophosphate, 8-azido-adenosine 3',5'-[32P]monophosphate (8-N3-cyclic AMP), was used to label the cyclic AMP binding proteins of Dictyostelium discoideum. During development cytosolic proteins appear which are specifically labeled by the photoaffinity agent. The proteins are developmentally regulated since they are only found in starve...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Proceedings of the National Academy of Sciences of the United States of America
دوره 67 1 شماره
صفحات -
تاریخ انتشار 1970