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B6db references: 9323022

type Journal Article
authors Wahl, M. C.; Huber, R.; Prade, L.; Marinkovic, S.; Messerschmidt, A.; Clausen, T.
title Cloning, purification, crystallization, and preliminary X-ray diffraction analysis of cystathionine gamma-synthase from E. coli
journal FEBS Lett
sel selected
ui 9323022
year (1997)
volume 414
number 3
pages 492-6
keywords Carbon-Oxygen Lyases/*chemistry/*genetics/metabolism
abstract The Escherichia coli metB gene has been PCR-extracted from genomic DNA and placed under the control of a tac and a T7 promoter in plasmids pCYB1 and pET22b(+), respectively, to produce overexpressing bacterial strains for the gene product, cystathionine gamma-synthase. Efficient purification procedures have been developed for a C-terminally intein- tagged version and the wild-type target protein, yielding the product in a quantity and homogeneity amenable to high-resolution single- crystal X-ray analysis. Crystals have been obtained in space group P1 with unit cell constants a=82.2 A, b=84.2 A, c=116.2 A, alpha=107.0 degrees, beta=96.3 degrees, gamma=108.0 degrees, suggesting eight monomers per asymmetric unit (V[M]=2.23 A3/Da). Crystals diffract to beyond 2.6 A resolution and a data set complete to 2.8 A resolution has been collected using a rotating anode X-ray source. A cryogenic buffer system has been developed to allow synchrotron data collection. Patterson self rotation searches reveal the presence of two independent tetramers with local 222 symmetry in an asymmetric unit. The crystallographic results corroborate and extend previous solution studies regarding the quaternary organization of the enzyme.
last changed 2009/02/11 18:28

B6db references