Crystallization and preliminary X-ray analysis of recombinant glutamate mutase and of the isolated component S from Clostridium cochlearium.
Glutamate mutase [varepsilon2sigma2(B12)1] was reconstituted by incubating purified components E (varepsilon2) and S (sigma2) from Clostridium cochlearium, both produced in Escherichia coli, with either aquo- or cyanocobalamin. The inactive glutamate mutase obtained was crystallized with polyethyleneglycol 4000 as precipitant. Crystals are monoclinic with space group P21 and have cell dimensions a = 64.6, b = 113.2, c = 108.4 A and beta = 96.0 degrees for the glutamate mutase reconstituted with aquocobalamin. They diffract to a resolution of at least 2.7 A. Isolated component S was crystallized in the presence of an excess of cyanocobalamin, yielding red crystals of space group I422 with unit-cell dimensions of a = b = 69.9 and c = 107.1 A. The crystals diffract to about 3.2 A resolution. Native data sets were collected for both crystal forms.
Department of Structural Biology, Institute of Physical Chemistry, University of Graz, Heinrichstrasse 28, 8010 Graz, Austria.
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Reitzer
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Krasser
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Jogl
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Buckel
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Bothe
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Kratky
D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D003013:Clostridium / Q000201:enzymology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D019751:Intramolecular Transferases / Q000737:chemistry* / Q000302:isolation & purification; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification
D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D019751:Intramolecular Transferases; C021570:methylaspartate mutase