3E0V

Crystal structure of pyruvate kinase from Leishmania mexicana in complex with sulphate ions

Summary for 3E0V

• Entry DOI: 10.2210/pdb3e0v/pdb
• Related: 3E0W
• Descriptor: Pyruvate kinase, SULFATE ION, GLYCEROL, ... (4 entities in total)
• Functional Keywords: pyruvate, kinase, nad+ nadh, adp, phosphoenolpyruvate, pep, glycolysis, trypanosomatid, leishmania, mexicana, allosteric enzyme, magnesium, metal-binding, transferase
• Biological source: Leishmania mexicana
• Total number of polymer chains: 6
• Total formula weight: 357325.42

Authors

Tulloch, L.B.,Gillmore, L.A.,Walkinshaw, M.D. (deposition date: 2008-08-01, release date: 2008-08-19, Last modification date: 2023-08-30)

Primary citation

Tulloch, L.B.,Morgan, H.P.,Hannaert, V.,Michels, P.A.,Fothergill-Gilmore, L.A.,Walkinshaw, M.D.
Sulphate removal induces a major conformational change in Leishmania mexicana pyruvate kinase in the crystalline state.
J.Mol.Biol., 383:615-626, 2008

PubMed Abstract: We report X-ray structures of pyruvate kinase from Leishmania mexicana (LmPYK) that are trapped in different conformations. These, together with the previously reported structure of LmPYK in its inactive (T-state) conformation, allow comparisons of three different conformers of the same species of pyruvate kinase (PYK). Four new site point mutants showing the effects of side-chain alteration at subunit interfaces are also enzymatically characterised. The LmPYK tetramer crystals grown with ammonium sulphate as precipitant adopt an active-like conformation, with sulphate ions at the active and effector sites. The sulphates occupy positions similar to those of the phosphates of ligands bound to active (R-state) and constitutively active (nonallosteric) PYKs from several species, and provide insight into the structural roles of the phosphates of the substrates and effectors. Crystal soaking in sulphate-free buffers was found to induce major conformational changes in the tetramer. In particular, the unwinding of the Aalpha6' helix and the inward hinge movement of the B domain are coupled with a significant widening (4 A) of the tetramer caused by lateral movement of the C domains. The two new LmPYK structures and the activity studies of site point mutations described in this article are consistent with a developing picture of allosteric activity in which localised changes in protein flexibility govern the distribution of conformer families adopted by the tetramer in its active and inactive states.

PubMed: 18775437
DOI: 10.1016/j.jmb.2008.08.037

Experimental method

X-RAY DIFFRACTION (3.3 Å)