Preparation and crystallization of a complex between human adenovirus serotype 2 proteinase and its 11-amino-acid cofactor pVIc.
Crystals have been obtained of the recombinant human adenovirus serotype 2 proteinase (AVP) in a complex with its 11-amino-acid cofactor pVIc. AVP-pVIc complexes were formed by the incubation of AVP with a 1.2-fold molar excess of pVIc prior to the crystallization trials. Diffraction-quality crystals were obtained at 18 degrees C by the vapor-diffusion method with 5.6 mg/ml AVP-pVIc in 1.4 M sodium acetate and 0.1 M Hepes, pH 7.5. Diffraction data (99% complete to 2.6 A resolution with Rmerge of 0.077) were collected from native crystals at room temperature at beamline X12-C at the National Synchrotron Light Source. The crystals belong to space group P6(1) with unit cell dimensions a = b = 114.2 A, c = 50.1 A; alpha = beta = 90 degrees, gamma = 120 degrees. The unit cell dimensions and likely mass of the molecular species in the crystals were consistent with there being one 25,000-Da complex (1:1) per asymmetric unit. Additionally, one heavy-atom derivative, obtained by the soaking of preformed crystals, was isomorphous to the native crystal. Diffraction data obtained on these crystals were 95% complete to 3.0 A resolution with an Rmerge of 0.076. Difference-Patterson analysis indicates three heavy atom sites in the derivative asymmetric unit.
McGrath
W J
WJ
Biology Department, Brookhaven National Laboratory, Upton, New York 11973, USA.
Ding
J
J
Sweet
R M
RM
Mangel
W F
WF
D003460:Crystallization; D018360:Crystallography, X-Ray; D003546:Cysteine Endopeptidases / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism
D011994:Recombinant Proteins; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; C091827:adenain