An improved strategy for the crystallization of Leishmania mexicana pyruvate kinase.
The inclusion of novel small molecules in crystallization experiments has provided very encouraging results and this method is now emerging as a promising alternative strategy for crystallizing 'problematic' biological macromolecules. These small molecules have the ability to promote lattice formation through stabilizing intermolecular interactions in protein crystals. Here, the use of 1,3,6,8-pyrenetetrasulfonic acid (PTS), which provides a helpful intermolecular bridge between Leishmania mexicana PYK (LmPYK) macromolecules in the crystal, is reported, resulting in the rapid formation of a more stable crystal lattice at neutral pH and greatly improved X-ray diffraction results. The refined structure of the LmPYK-PTS complex revealed the negatively charged PTS molecule to be stacked between positively charged (surface-exposed) arginine side chains from neighbouring LmPYK molecules in the crystal lattice.
Morgan
Hugh P
HP
Structural Biochemistry Group, Institute of Structural and Molecular Biology, The University of Edinburgh, Michael Swann Building, The King's Buildings, Mayfield Road, Edinburgh EH9 3JR, Scotland.
McNae
Iain W
IW
Hsin
Kun-Yi
KY
Michels
Paul A M
PA
Fothergill-Gilmore
Linda A
LA
Walkinshaw
Malcolm D
MD
D018360:Crystallography, X-Ray; D007894:Leishmania mexicana / Q000201:enzymology*; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D011770:Pyruvate Kinase / Q000737:chemistry* / Q000378:metabolism; D013379:Substrate Specificity; D013451:Sulfonic Acids / Q000737:chemistry / Q000378:metabolism
D013451:Sulfonic Acids; D011770:Pyruvate Kinase