Crystallization and preliminary X-ray analysis of Streptococcus pneumoniae hyaluronate lyase.
A fully active 83-kDa truncated form of recombinant hyaluronate lyase from Streptococcus pneumoniae was crystallized by the hanging drop vapor diffusion method using ammonium sulfate as a precipitating agent. Crystals grew at room temperature using a variety of buffers with pH around 6. The crystals diffract X-rays beyond 2.0 A resolution using Cu K alpha radiation and a rotating-anode X-ray source. They belong to the orthorhombic space group P2(1)2(1)2(1) with unit cell dimensions, a = 84.2, b = 104.2, c = 104.6 A, and alpha = beta = gamma = 90.0 degrees. The VM value of 2.9 A3/Da is consistent with only one molecule of the enzyme in the asymmetric unit and the solvent content of 57%. Diffraction data 94.7% complete to 2.0 A resolution with Rsym of 5.4% were collected from one native crystal at room temperature. The search for heavy-atom derivatives to solve the structure is in progress.
Jedrzejas
M J
MJ
Department of Microbiology, University of Alabama at Birmingham 35294, USA.
Chantalat
L
L
Mewbourne
R B
RB
D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011133:Polysaccharide-Lyases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013296:Streptococcus pneumoniae / Q000201:enzymology* / Q000235:genetics
D010446:Peptide Fragments; D011994:Recombinant Proteins; D011133:Polysaccharide-Lyases; C100002:hyaluronate lyase