Molecular properties of the putative nitrogen sensor PII from Arabidopsis thaliana.
Although the signal sensing protein PII is well known to play a central role in bacterial nitrogen metabolism, the structure and function of PII in plants remains only partially understood. Comparative modeling was undertaken based on the high degree of amino acid identity between Escherichia coli and Arabidopsis PII. The mature Arabidopsis PII predicted structure superimposes very well onto the E. coli PII structure (Calpha root mean square deviation < 0.4 A). The model of the highly conserved T-loop suggests a molecular mechanism by which the plant PII may regulate putative post-translational modification in response to metabolite binding. Consistent with the presence of key conserved residues necessary for trimer formation, gel filtration showed the oligomeric structure of Arabidopsis thaliana PII to be a homotrimer. We have demonstrated that Arabidopsis PII binds to the small molecules, ATP, ADP, 2KG, and with lesser affinity to OAA, using isothermal titration calorimetry. We have determined the metabolite dissociation constants and compared these with known physiological concentrations of these metabolites in the plant to identify the Arabidopsis PII effector molecules and their possible roles. We predict that the plant PII is likely continually bound by ATP, and its ligand-bound state only varying with respect to the degree of 2KG binding. Based on our in vitro binding studies, the function of plant PII as a 2KG sensor is suggested.
Smith
Catherine S
CS
Department of Biological Sciences, University of Calgary, 2500 University Drive NW, Calgary, Alberta, Canada T2N 1N4.
Weljie
Aalim M
AM
Moorhead
Greg B G
GB
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D017360:Arabidopsis / Q000737:chemistry / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000378:metabolism*; D002151:Calorimetry; D008024:Ligands; D008958:Models, Molecular; D009584:Nitrogen / Q000378:metabolism*; D051922:PII Nitrogen Regulatory Proteins; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D013816:Thermodynamics
D029681:Arabidopsis Proteins; D008024:Ligands; D051922:PII Nitrogen Regulatory Proteins; C473074:PII protein, Arabidopsis; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D009584:Nitrogen