Binding constant determination studies utilizing recombinant delta CREB protein.
In this study, we report the binding constants (Kd) of the cAMP-responsive element binding protein (delta-CREB) for various cAMP-response element (CRE) motifs. We utilized purified recombinant delta CREB protein in binding reactions with natural CRE motifs found in the promoter of two neuropeptide hormone genes and with several variant CRE motifs. The Kd of delta CREB for the perfectly palindromic CRE, TGACG-TCA, found within the somatostatin promoter is estimated to be 5.0 x 10(-9) M. The Kd of delta CREB for the variant CRE motif TG_CGTCA found within the enkephalin promoter is calculated to be in the 3 x 10(-8) M. These studies provide an in vitro quantitative assessment of the binding affinity of delta CREB for various CRE motifs.
Williams
J S
JS
Department of Veterinary Physiology and Pharmacology, Purdue University, W. Lafayette, IN 47907-1246.
Dixon
J E
JE
Andrisani
O M
OM
D000595:Amino Acid Sequence; D001483:Base Sequence; D017362:Cyclic AMP Response Element-Binding Protein / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D011485:Protein Binding; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000502:physiology*
D017362:Cyclic AMP Response Element-Binding Protein; D011994:Recombinant Proteins