Characterization of caged compounds binding to proteins by NMR spectroscopy.
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.
Bandorowicz-Pikula
Joanna
J
Department of Biochemistry, Nencki Institute of Experimental Biology, 3 Pasteur Street, 02093 Warsaw, Poland.
Buchet
René
R
Cañada
F Javier
FJ
Clémancey
Martin
M
Groves
Patrick
P
Jiménez-Barbero
Jesus
J
Lancelin
Jean-Marc
JM
Marcillat
Olivier
O
Pikula
Slawomir
S
Sekrecka-Belniak
Anna
A
Strzelecka-Kiliszek
Agnieszka
A
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
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