1TUY

Acetate Kinase complexed with ADP, AlF3 and acetate

Summary for 1TUY

• Entry DOI: 10.2210/pdb1tuy/pdb
• Related: 1TUU
• Descriptor: Acetate kinase, SULFATE ION, ADENOSINE-5'-DIPHOSPHATE, ... (6 entities in total)
• Functional Keywords: alpha/beta, askha (acetate and sugar kinases, hsc70, actin) superfamily, alf3 and acetate, transferase
• Biological source: Methanosarcina thermophila
• Cellular location: Cytoplasm (By similarity): P38502
• Total number of polymer chains: 2
• Total formula weight: 87918.03

Authors

Gorrell, A.,Lawrence, S.H.,Ferry, J.G. (deposition date: 2004-06-25, release date: 2005-01-11, Last modification date: 2024-02-14)

Primary citation

Gorrell, A.,Lawrence, S.H.,Ferry, J.G.
Structural and Kinetic Analyses of Arginine Residues in the Active Site of the Acetate Kinase from Methanosarcina thermophila.
J.Biol.Chem., 280:10731-10742, 2005

PubMed Abstract: Acetate kinase catalyzes transfer of the gamma-phosphate of ATP to acetate. The only crystal structure reported for acetate kinase is the homodimeric enzyme from Methanosarcina thermophila containing ADP and sulfate in the active site (Buss, K. A., Cooper, D. C., Ingram-Smith, C., Ferry, J. G., Sanders, D. A., and Hasson, M. S. (2001) J. Bacteriol. 193, 680-686). Here we report two new crystal structure of the M. thermophila enzyme in the presence of substrate and transition state analogs. The enzyme co-crystallized with the ATP analog adenosine 5'-[gamma-thio]triphosphate contained AMP adjacent to thiopyrophosphate in the active site cleft of monomer B. The enzyme co-crystallized with ADP, acetate, Al(3+), and F(-) contained a linear array of ADP-AlF(3)-acetate in the active site cleft of monomer B. Together, the structures clarify the substrate binding sites and support a direct in-line transfer mechanism in which AlF(3) mimics the meta-phosphate transition state. Monomers A of both structures contained ADP and sulfate, and the active site clefts were closed less than in monomers B, suggesting that domain movement contributes to catalysis. The finding that His(180) was in close proximity to AlF(3) is consistent with a role for stabilization of the meta-phosphate that is in agreement with a previous report indicating that this residue is essential for catalysis. Residue Arg(241) was also found adjacent to AlF(3), consistent with a role for stabilization of the transition state. Kinetic analyses of Arg(241) and Arg(91) replacement variants indicated that these residues are essential for catalysis and also indicated a role in binding acetate.

PubMed: 15647264
DOI: 10.1074/jbc.M412118200

Experimental method

X-RAY DIFFRACTION (3 Å)