title: Characterization of caged compounds binding to proteins by NMR spectroscopy.
abstract: Photolysable caged ligands are used to investigate protein function and activity. Here, we investigate the binding properties of caged nucleotides and their photo released products to well established but evolutionary and structurally unrelated nucleotide-binding proteins, rabbit muscle creatine kinase (RMCK) and human annexin A6 (hAnxA6), using saturation transfer difference NMR spectroscopy. We detect the binding of the caged nucleotides and discuss the general implications on interpreting data collected with photolysable caged ligands using different techniques. Strategies to avoid non-specific binding of caged compound to certain proteins are also suggested.
authors.affiliation: Department of Biochemistry, Nencki Institute of Experimental Biology, 3 Pasteur Street, 02093 Warsaw, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Bandorowicz-Pikula
authors.forename: René
authors.initials: R
authors.lastname: Buchet
authors.forename: F Javier
authors.initials: FJ
authors.lastname: Cañada
authors.forename: Martin
authors.initials: M
authors.lastname: Clémancey
authors.forename: Patrick
authors.initials: P
authors.lastname: Groves
authors.forename: Jesus
authors.initials: J
authors.lastname: Jiménez-Barbero
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Lancelin
authors.forename: Olivier
authors.initials: O
authors.lastname: Marcillat
authors.forename: Slawomir
authors.initials: S
authors.lastname: Pikula
authors.forename: Anna
authors.initials: A
authors.lastname: Sekrecka-Belniak
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Strzelecka-Kiliszek
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D017317:Annexin A6 / Q000737:chemistry / Q000378:metabolism*; D052283:Creatine Kinase, MM Form / Q000737:chemistry / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008024:Ligands; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011485:Protein Binding; D011817:Rabbits
chemical_list: D017317:Annexin A6; D008024:Ligands; C552741:guanosine 5'-O-(3-thiotriphosphate) P-3-(1-(4,5-dimethoxy-2-nitrophenyl)ethyl) ester; C029273:P(3)-1-(2-nitro)phenylethyladenosine 5'-triphosphate; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D052283:Creatine Kinase, MM Form