Acta Crystallographica Section F

Structural Biology and Crystallization Communications

Volume 64, Part 3 (March 2008)

crystallization communications

Acta Cryst. (2008). F64, 179-181 [ doi:10.1107/S1744309108003473 ]

Crystallization and preliminary X-ray diffraction analysis of full-length and proteolytically activated pyruvate oxidase from Escherichia coli

A. Weidner, P. Neumann, G. Wille, M. T. Stubbs and K. Tittmann

Abstract: The thiamine diphosphate- and flavin-dependent peripheral membrane enzyme pyruvate oxidase from Escherichia coli (EcPOX) has been crystallized in the full-length form and as a proteolytically activated C-terminal truncation variant which lacks the last 23 amino acids ( [Delta] 23 EcPOX). Crystals were grown by the hanging-drop vapour-diffusion method using either protamine sulfate (full-length EcPOX) or 2-methyl-2,4-pentanediol ( [Delta] 23 EcPOX) as precipitants. Native data sets were collected at a X-ray home source to a resolution of 2.9 Å. The two forms of EcPOX crystallize in different space groups. Whereas full-length EcPOX crystallizes in the tetragonal space group P4₃2₁2 with two monomers per asymmetric unit, the crystals of [Delta] 23 EcPOX belong to the orthorhombic space group P2₁2₁2₁ and contain 12 monomers per asymmetric unit.

Keywords: peripheral membrane proteins; thiamine diphosphate; flavin adenine dinucleotide; ubiquinone; pyruvate; electron transfer; decarboxylation.