2AN9

Crystal Structure Of Oligomeric E.coli Guanylate Kinase In Complex With GDP

2AN9 の概要

• エントリーDOI: 10.2210/pdb2an9/pdb
• 関連するPDBエントリー: 2ANB 2ANC
• 分子名称: Guanylate kinase, POTASSIUM ION, GUANOSINE-5'-DIPHOSPHATE, ... (5 entities in total)
• 機能のキーワード: transferase, gmp kinase, guanylate kinase, oligomeric
• 由来する生物種: Escherichia coli
• 細胞内の位置: Cytoplasm: P60546
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 48782.53

構造登録者

Hible, G.,Renault, L.,Schaeffer, F.,Christova, P.,Radulescu, A.Z.,Evrin, C.,Gilles, A.M.,Cherfils, J. (登録日: 2005-08-11, 公開日: 2005-08-30, 最終更新日: 2023-10-25)

主引用文献

Hible, G.,Renault, L.,Schaeffer, F.,Christova, P.,Radulescu, A.Z.,Evrin, C.,Gilles, A.M.,Cherfils, J.
Calorimetric and crystallographic analysis of the oligomeric structure of Escherichia coli GMP kinase
J.Mol.Biol., 352:1044-1059, 2005

PubMed Abstract: Guanosine monophosphate kinases (GMPKs), which catalyze the phosphorylation of GMP and dGMP to their diphosphate form, have been characterized as monomeric enzymes in eukaryotes and prokaryotes. Here, we report that GMPK from Escherichia coli (ecGMPK) assembles in solution and in the crystal as several different oligomers. Thermodynamic analysis of ecGMPK using differential scanning calorimetry shows that the enzyme is in equilibrium between a dimer and higher order oligomers, whose relative amounts depend on protein concentration, ionic strength, and the presence of ATP. Crystallographic structures of ecGMPK in the apo, GMP and GDP-bound forms were solved at 3.2A, 2.9A and 2.4A resolution, respectively. ecGMPK forms a hexamer with D3 symmetry in all crystal forms, in which the two nucleotide-binding domains are able to undergo closure comparable to that of monomeric GMPKs. The 2-fold and 3-fold interfaces involve a 20-residue C-terminal extension and a sequence signature, respectively, that are missing from monomeric eukaryotic GMPKs, explaining why ecGMPK forms oligomers. These signatures are found in GMPKs from proteobacteria, some of which are human pathogens. GMPKs from these bacteria are thus likely to form the same quaternary structures. The shift of the thermodynamic equilibrium towards the dimer at low ecGMPK concentration together with the observation that inter-subunit interactions partially occlude the ATP-binding site in the hexameric structure suggest that the dimer may be the active species at physiological enzyme concentration.

PubMed: 16140325
DOI: 10.1016/j.jmb.2005.07.042

実験手法

X-RAY DIFFRACTION (2.35 Å)