Cyclic nucleotide binding proteins in the Arabidopsis thaliana and Oryza sativa genomes - Figure 2

<p><strong>Copyright information:</strong></p> <p>Taken from "Cyclic nucleotide binding proteins in the and genomes"</p> <p>BMC Bioinformatics 2005;6():6-6.</p> <p>Published online 11 Jan 2005</p> <p>PMCID:PMC545951.</p> <p>Copyright © 2005 Bridges et al; licensee BioMed Central Ltd.</p> <p>re aligned against several well studied CNB domains including regulatory subunits of PKA (RIα and RIIβ), Epac1, Epac2, and cyclic GMP dependent kinase 2 (CGK2) from humans, HCN2 from mouse and CAP. Highlighted on the alignment are glycine residues involved in loop structures (dark grey arrows), residues forming the hydrophobic pocket for cNMP binding (green arrows) and residues proposed to contact the phosphate of the cNMP (blue arrows). The highly conserved helix capping acidic residue is shown in red. Secondary structure is denoted by arrows above the alignment, with light blue for alpha helices and pink for beta sheets and is based on the secondary structure of HCN2. (B) A homology model of atCNTE1 was generated from the known structures of CNB domains. Key residues are shown as stick representations and are colored and labeled according to the color scheme described in (A). The cGMP ligand is shown in magenta and is based on the structure of cGMP bound to HCN2 [pdb: 1Q3E] superimposed over our model. Figure was generated with Molscript [83] and Raster3D [84].</p

Cyclic nucleotide binding proteins in the Arabidopsis thaliana and Oryza sativa genomes - Figure 3

<p><strong>Copyright information:</strong></p> <p>Taken from "Cyclic nucleotide binding proteins in the and genomes"</p> <p>BMC Bioinformatics 2005;6():6-6.</p> <p>Published online 11 Jan 2005</p> <p>PMCID:PMC545951.</p> <p>Copyright © 2005 Bridges et al; licensee BioMed Central Ltd.</p> <p>re aligned against several well studied CNB domains including regulatory subunits of PKA (RIα and RIIβ), Epac1, Epac2, and cyclic GMP dependent kinase 2 (CGK2) from humans, HCN2 from mouse and CAP. Highlighted on the alignment are glycine residues involved in loop structures (dark grey arrows), residues forming the hydrophobic pocket for cNMP binding (green arrows) and residues proposed to contact the phosphate of the cNMP (blue arrows). The highly conserved helix capping acidic residue is shown in red. Secondary structure is denoted by arrows above the alignment, with light blue for alpha helices and pink for beta sheets and is based on the secondary structure of HCN2. (B) A homology model of atCNTE1 was generated from the known structures of CNB domains. Key residues are shown as stick representations and are colored and labeled according to the color scheme described in (A). The cGMP ligand is shown in magenta and is based on the structure of cGMP bound to HCN2 [pdb: 1Q3E] superimposed over our model. Figure was generated with Molscript [83] and Raster3D [84].</p

Cyclic nucleotide binding proteins in the Arabidopsis thaliana and Oryza sativa genomes - Figure 4

<p><strong>Copyright information:</strong></p> <p>Taken from "Cyclic nucleotide binding proteins in the <em>Arabidopsis thaliana</em> and <em>Oryza sativa</em> genomes"</p> <p>BMC Bioinformatics 2005;6():6-6.</p> <p>Published online 11 Jan 2005</p> <p>PMCID:PMC545951.</p> <p>Copyright © 2005 Bridges et al; licensee BioMed Central Ltd.</p> <p><strong>Biochemical evidence for lack of a cyclic nucleotide dependent kinase in <em>Arabidopsis thaliana</em></strong>. (A) Protein kinase assays using Kemptide as a substrate. Assays were conducted on identically prepared extracts of Arabidopsis and rat adipose tissue in the presence or absence (control) of 10 μM cyclic nucleotide as indicated. Scale is offset in order to visualize both sets of results. All assays were performed in duplicate from three separate preparations and error bars indicate standard error for three separate preparations. (B) Western blotting of extracts with PKA catalytic (PKAcs) and regulatory (RII) subunit polyclonal antibodies. The PKAcs antibody was affinity purified according to [82] and used at 0.5 μg/mL while the RII antibody was used as crude serum at 5000X dilution. Lanes are as follows (A), 10 ng of purified bovine PKAcs or RII, (B) 25 μg clarified crude Arabidopsis extract, (C) 25 μg clarified crude rat adipocyte extract. Positions of mammalian PKA and RII are indicated with arrows.</p