Open AccessCrystallization and preliminary X‐ray diffraction analysis of phosphoglycerate kinase from Streptococcus pneumoniae
Author(s) -
BernardoGarcía Noelia,
Bartual Sergio G.,
Fulde Marcus,
Bergmann Simone,
Hermoso Juan A.
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111030922
Subject(s) - phosphoglycerate kinase , crystallization , monoclinic crystal system , crystallography , glycolysis , biology , enzyme , chemistry , biochemistry , crystal structure , organic chemistry
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 I 222, 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 I 222 crystals. The ternary complex of PGK with adenylyl imidodiphosphate (AMP‐PNP) and 3‐phospho‐ d ‐glycerate (3PGA) produced monoclinic crystals (space group P 2 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.