Structures of adenylosuccinate synthetase from Triticum aestivum and Arabidopsis thaliana.
Catalyzing the first step in the de novo synthesis of adenylmonophosphate, adenylosuccinate synthetase (AdSS) is a known target for herbicides and antibiotics. We have purified and crystallized recombinant AdSS from Arabidopsis thaliana and Tritium aestivum, expressed in Escherichia coli. The structures of A. thaliana and T. aestivum AdSS in complex with GDP were solved at 2.9 A and 3.0 A resolution, respectively. Comparison with the known structures from E. coli reveals that the overall fold is very similar to that of the E. coli protein. The longer N terminus in the plant sequences is at the same place as the longer C terminus of the E. coli sequence in the 3D structure. The GDP-binding sites have one additional hydrogen-bonding partner, which is a plausible explanation for the lower K(m) value. Due to its special position, this partner may also enable GTP to initiate a conformational change, which was, in E. coli AdSS, exclusively activated by ligands at the IMP-binding site. The dimer interfaces show up to six hydrogen bonds and six salt-bridges more than in the E. coli structure, although the contact areas have approximately the same size.
Prade
L
L
Novartis Pharma AG, Basel, CH-4056, Switzerland.
Cowan-Jacob
S W
SW
Chemla
P
P
Potter
S
S
Ward
E
E
Fonne-Pfister
R
R
D000265:Adenylosuccinate Synthase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology*; D001665:Binding Sites; D018360:Crystallography, X-Ray; D019281:Dimerization; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D006153:Guanosine Diphosphate / Q000378:metabolism*; D006860:Hydrogen Bonding; D007291:Inosine Monophosphate / Q000378:metabolism; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D055672:Static Electricity; D014908:Triticum / Q000201:enzymology*
D008024:Ligands; D011994:Recombinant Proteins; D007291:Inosine Monophosphate; D006153:Guanosine Diphosphate; D000265:Adenylosuccinate Synthase