Crystallization of the pneumococcal autolysin LytC: in-house phasing using novel lanthanide complexes.
LytC, one of the major autolysins from the human pathogen Streptococcus pneumoniae, has been crystallized as needles by the hanging-drop technique using 10%(w/v) PEG 3350 as precipitant and 10 mM HEPES pH 7.5. LytC crystals were quickly soaked in mother liquor containing 2 mM of the complex Gd-HPDO3A to produce derivatized crystals (LytC(Gd-HPDO3A)). Both native LytC and isomorphous LytC(Gd-HPDO3A) crystals were flash-cooled in a nitrogen flow at 120 K prior to X-ray data collection using an in-house Enraf-Nonius rotating-anode generator (lambda = 1.5418 A) and a MAR345 imaging-plate detector. In both cases, good-quality diffraction patterns were obtained at high resolution. LytC(Gd-HPDO3A) crystals allowed the collection of a SAD X-ray data set to 2.6 A resolution indexed in terms of a P2(1) monoclinic unit cell with parameters a = 59.37, b = 67.16, c = 78.85 A, beta = 105.69 degrees . The anomalous Patterson map allowed the identification of one heavy-atom binding site, which was sufficient for the calculation of an interpretable anomalous map at 2.6 A resolution.
Pérez-Dorado
Inmaculada
I
Grupo de Cristalografía Macromolecular y Biología Estructural, Instituto de Química-Física Rocasolano, CSIC, Serrano 119, 28006 Madrid, Spain.
Sanles
Reyes
R
González
Ana
A
García
Pedro
P
García
José L
JL
Martínez-Ripoll
Martín
M
Hermoso
Juan A
JA
D003460:Crystallization; D018360:Crystallography, X-Ray; D005682:Gadolinium; D006571:Heterocyclic Compounds / Q000737:chemistry*; D015394:Molecular Structure; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000737:chemistry*; D009942:Organometallic Compounds / Q000737:chemistry*; D013296:Streptococcus pneumoniae / Q000201:enzymology*
D006571:Heterocyclic Compounds; D009942:Organometallic Compounds; C062402:gadoteridol; D005682:Gadolinium; C419222:LytC protein, Streptococcus; D009238:N-Acetylmuramoyl-L-alanine Amidase
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