Crystallization and Initial X-Ray Diffraction Analysis of Human Pyruvate Dehydrogenase

Abstract

Human pyruvate dehydrogenase (E1) is a component enzyme of the pyruvate dehydrogenase complex. The enzyme catalyzes the decarboxylation of pyruvate followed by a reductive acetylation of lipoyl groups of the dihydrolipoamide acetyltransferase component of the pyruvate dehydrogenase complex. El is an alpha(sub 2)Beta(sub 2) tetrameric assembly of an approximate molecular mass of 154 kDa. The crystals of this recombinant enzyme have been grown from polyethylene glycol 3350 using vapor diffusion method at 295K. The crystals are characterized as orthorhombic, space group P2(sub 1)2(sub 1)2(sub 1), with cell parameters of a = 64.2, b = 126.9 and c = 190.2 A. Crystals diffracted to a minimum d-spacing of 2.5 A. The asymmetric unit contains one alpha(sub 2)Beta(sub 2) tetrameric El assembly, and self-rotation function analysis showed a pseudo-twofold symmetry relating the two monomers.