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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
25
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pubmed:dateCreated |
1990-10-9
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pubmed:abstractText |
X-ray crystallographic diffraction data has been collected for recombinant hexadecameric ribulose-P2 carboxylase from the cyanobacterium Synechococcus PCC6301 expressed in Escherichia coli. The enzyme has been purified and then crystallized in a number of crystal forms from polyethylene glycol solutions. The best crystals were obtained with enzyme that was first activated with the cofactors CO2 and Mg2+ in the presence of the tight-binding intermediate analogue, 2'-carboxyarabinitol 1,5-bisphosphate. One crystal form with plate-like morphology diffracts beyond 2.5 A but has one axis greater than 350 A. A second crystal form that diffracts to similar resolution grows with space group P212121 and unit cell dimensions of a = 223.9 A, b = 111.9 A, and c = 199.7 A. The crystal forms used to collect the diffraction data have been redissolved to determine that the recombinant ribulose-P2 carboxylase L8S8 molecule is indeed composed of equal numbers of large and small subunits and also that a quaternary complex between activated ribulose-P2 carboxylase E.CO2.Mg2+, and the analogue was present in the crystals. Denaturation of the redissolved enzyme in the absence of thiol-reducing agents established that the L-subunits of the L8 core are substantially dimeric, cross-linked by a disulfide bridge. Crystals of spinach ribulose-P2 carboxylase were likewise analyzed to show that dimers of the L-subunit were also predominant. This report identifies a single cysteine residue in the L-subunit that forms a bridge between those L-monomers that compose the four putative functional dimers of the L8 core.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Sep
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pubmed:issn |
0021-9258
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
5
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pubmed:volume |
265
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
15154-9
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pubmed:dateRevised |
2006-5-1
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pubmed:meshHeading |
pubmed-meshheading:2118521-Chromatography, Ion Exchange,
pubmed-meshheading:2118521-Crystallization,
pubmed-meshheading:2118521-Cyanobacteria,
pubmed-meshheading:2118521-Enzyme Activation,
pubmed-meshheading:2118521-Escherichia coli,
pubmed-meshheading:2118521-Macromolecular Substances,
pubmed-meshheading:2118521-Models, Molecular,
pubmed-meshheading:2118521-Plasmids,
pubmed-meshheading:2118521-Protein Conformation,
pubmed-meshheading:2118521-Recombinant Proteins,
pubmed-meshheading:2118521-Ribulose-Bisphosphate Carboxylase,
pubmed-meshheading:2118521-X-Ray Diffraction
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pubmed:year |
1990
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pubmed:articleTitle |
The purification and preliminary X-ray diffraction studies of recombinant Synechococcus ribulose-1,5-bisphosphate carboxylase/oxygenase from Escherichia coli.
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pubmed:affiliation |
Department of Molecular Biology, Uppsala Biomedical Centre, Sweden.
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pubmed:publicationType |
Journal Article
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