Download MendeleyCrystal structures of mutant monomeric hexokinase I reveal multiple ADP binding sites and conformational changes relevant to allosteric regulation.
Aleshin, A.E., Kirby, C., Liu, X., Bourenkov, G.P., Bartunik, H.D., Fromm, H.J., Honzatko, R.B.(2000) J Mol Biol 296: 1001-1015
- PubMed: 10686099
- DOI: https://doi.org/10.1006/jmbi.1999.3494
- Primary Citation of Related Structures:
1CZA, 1DGK - PubMed Abstract:
Hexokinase I, the pacemaker of glycolysis in brain tissue, is composed of two structurally similar halves connected by an alpha-helix. The enzyme dimerizes at elevated protein concentrations in solution and in crystal structures; however, almost all published data reflect the properties of a hexokinase I monomer in solution. Crystal structures of mutant forms of recombinant human hexokinase I, presented here, reveal the enzyme monomer for the first time. The mutant hexokinases bind both glucose 6-phosphate and glucose with high affinity to their N and C-terminal halves, and ADP, also with high affinity, to a site near the N terminus of the polypeptide chain. Exposure of the monomer crystals to ADP in the complete absence of glucose 6-phosphate reveals a second binding site for adenine nucleotides at the putative active site (C-half), with conformational changes extending 15 A to the contact interface between the N and C-halves. The structures reveal distinct conformational states for the C-half and a rigid-body rotation of the N-half, as possible elements of a structure-based mechanism for allosteric regulation of catalysis.
Organizational Affiliation:
Department of Biochemistry Biophysics and Molecular Biology, Iowa State University, Ames, IA 50011, USA.
