Expression, crystallization and preliminary X-ray diffraction studies of recombinant Clostridium perfringens beta 2-toxin.
Clostridium perfringens is a Gram-positive sporulating anaerobic bacterium that is responsible for a wide spectrum of diseases in animals, birds and humans. The virulence of C. perfringens is associated with the production of several enterotoxins and exotoxins. beta2-toxin is a 28 kDa exotoxin produced by C. perfringens. It is implicated in necrotic enteritis in animals and humans, a disease characterized by a sudden acute onset with lethal hemorrhagic mucosal ulceration. The recombinant expression, purification and crystallization of beta2-toxin using the batch-under-oil technique are reported here. Native X-ray diffraction data were obtained to 2.9 A resolution on a synchrotron beamline at the F2 station at Cornell High Energy Synchrotron Source (CHESS) using an ADSC Quantum-210 CCD detector. The crystals belong to space group R3, with a dimer in the asymmetric unit; the unit-cell parameters are a = b = 103.71, c = 193.48 A, alpha = beta = 90, gamma = 120 degrees using the hexagonal axis setting. A self-rotation function shows that the two molecules are related by a noncrystallographic twofold axis with polar angles omega = 90.0, phi = 210.3 degrees.
Gurjar
Abhijit A
AA
Department of Veterinary and Biomedical Science, The Pennsylvania State University, PA, USA.
Yennawar
Neela H
NH
Yennawar
Hemant P
HP
Rajashankar
Kanagalaghatta R
KR
Hegde
Narasimha V
NV
Jayarao
Bhushan M
BM
D001427:Bacterial Toxins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003016:Clostridium perfringens / Q000737:chemistry / Q000235:genetics; D003460:Crystallization; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D014961:X-Ray Diffraction / Q000379:methods*
D001427:Bacterial Toxins; D011994:Recombinant Proteins; C109896:cpb2 protein, Clostridium perfringens