title: Crystallization and preliminary X-ray diffraction analysis of phosphoglycerate kinase from Streptococcus pneumoniae.
abstract: Phosphoglycerate kinase (PGK) is a widespread two-domain enzyme that plays a critical role in the glycolytic pathway. Several glycolytic enzymes from streptococci have been identified as surface-exposed proteins that are involved in streptococcal virulence by their ability to bind host proteins. This binding allows pneumococcal cells to disseminate through the epithelial and endothelial layers. Crystallization of PGK from Streptococcus pneumoniae yielded orthorhombic crystals (space group I222, unit-cell parameters a = 62.73, b = 75.38, c = 83.63 Å). However, the unit cell of these crystals was not compatible with the presence of full-length PGK. Various analytical methods showed that only the N-terminal domain of PGK was present in the I222 crystals. The ternary complex of PGK with adenylyl imidodiphosphate (AMP-PNP) and 3-phospho-D-glycerate (3PGA) produced monoclinic crystals (space group P2(1), unit-cell parameters a = 40.35, b = 78.23, c = 59.03 Å, β = 96.34°). Molecular replacement showed that this new crystal form contained full-length PGK, thereby indicating the relevance of including substrates in order to avoid proteolysis during the crystallization process.
authors.affiliation: Department of Crystallography and Structural Biology, Instituto de Química-Física Rocasolano, CSIC, Serrano 119, 28006 Madrid, Spain.
authors.forename: Noelia
authors.initials: N
authors.lastname: Bernardo-García
authors.forename: Sergio G
authors.initials: SG
authors.lastname: Bartual
authors.forename: Marcus
authors.initials: M
authors.lastname: Fulde
authors.forename: Simone
authors.initials: S
authors.lastname: Bergmann
authors.forename: Juan A
authors.initials: JA
authors.lastname: Hermoso
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D010735:Phosphoglycerate Kinase / Q000737:chemistry*; D013296:Streptococcus pneumoniae / Q000201:enzymology*
chemical_list: D010735:Phosphoglycerate Kinase
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