C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/48 (2006.01) A61K 49/06 (2006.01) C12N 5/10 (2006.01) C12P 21/06 (2006.01)
Patent
CA 2547456
The present invention relates to a chimeric peptide, comprising a cAMP binding moiety having only one cAMP binding site and at least two detectable labels, whereby the first of said two detectable labels is located at the carboxy terminus and the second of said two detectable labels is located at the amino terminus of said cAMP binding moiety. Said chimeric peptide of the invention is particularly useful in/for direct determination of cAMP concentrations) in vitro and/or in vivo. Furthermore, nucleic acid molecules encoding said chimeric proteins are described as well as vectors and host cells comprising the same. The present invention also provides methods for producing the chimeric protein of the invention and methods for identification and screening of molecules or compounds which are capable of modifying cAMP binding to the chimeric peptide of the invention or the biological and/or pharmacological function of adenylyl cyclases or phosphodiesterases. In addition, a method for cAMP determination in a sample and a method for the detection of cAMP in the living cell or tissue is described. Finally, a kit comprising the compounds of the present invention is disclosed.
L'invention concerne un peptide chimère qui comprend un fragment de liaison cAMP ne comportant qu'un site de liaison cAMP, et au moins deux étiquettes détectables, la première des deux étiquettes détectables étant située sur la terminaison carboxy et la seconde des deux étiquettes détectables étant située sur la terminaison amino dudit fragment de liaison cAMP. Le peptide chimère de l'invention est particulièrement utile pour la mesure directe des concentrations de cAMP in vitro et/ou in vivo. L'invention porte en outre sur des molécules d'acides nucléiques codant lesdites protéines chimères, et sur des vecteurs et cellules hôtes renfermant ces dernières. L'invention se rapporte également à des procédés de production de la protéine chimère et à des procédés d'identification et de criblage de molécules ou de composés qui sont capables de modifier la liaison de cAMP au peptide chimère de l'invention ou la fonction biologique et/ou pharmacologique des adénylyl cyclases ou des phosphodiesterases. L'invention concerne en outre un procédé qui permet de mesurer cAMP dans un échantillon, et un procédé qui permet de détecter cAMP dans un tissu ou une cellule vivante. L'invention porte enfin sur une trousse comprenant les composés de l'invention.
Bunemann Moritz
Lohse Martin J.
Nikolaev Viacheslav
Bayerische Julius-Maximilians-Universitat Wurzburg
Goudreau Gage Dubuc
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