43 research outputs found
Recommended from our members
Preliminary analysis of crystals of satellite tobacco mosaic virus.
Satellite tobacco mosaic virus (STMV), a small T = 1 icosahedral plant virus, has been crystallized in a form suitable for high-resolution X-ray diffraction analysis. The crystals, which diffract to better than 2.5 A resolution, are of space group I222 and have unit cell dimensions of a = 176 A, b = 192 A and c = 205 A. The centers of the virus particles occupy 222 symmetry points in the unit cell and one quarter of the virus particle constitutes the asymmetric unit, which is therefore comprised of 15 capsid protein molecules. From packing considerations, the maximum diameter of the STMV particles cannot exceed 165 A, and it is probably 5 to 10 A less than this value
Recommended from our members
Preliminary analysis of crystals of satellite tobacco mosaic virus.
Satellite tobacco mosaic virus (STMV), a small T = 1 icosahedral plant virus, has been crystallized in a form suitable for high-resolution X-ray diffraction analysis. The crystals, which diffract to better than 2.5 A resolution, are of space group I222 and have unit cell dimensions of a = 176 A, b = 192 A and c = 205 A. The centers of the virus particles occupy 222 symmetry points in the unit cell and one quarter of the virus particle constitutes the asymmetric unit, which is therefore comprised of 15 capsid protein molecules. From packing considerations, the maximum diameter of the STMV particles cannot exceed 165 A, and it is probably 5 to 10 A less than this value
Recommended from our members
Determination of local refractive index for protein and virus crystals in solution by Mach-Zehnder interferometry.
To establish the importance of, and quantitatively evaluate, the macromolecular concentration gradients in the neighborhood of growing protein, virus, and nucleic acid crystals, a convenient, accurate, and nonintrusive method has been devised. This approach should prove particularly relevant in the rigorous comparison of crystals grown in a conventional laboratory setting with those grown in a microgravity environment. The method is based on precise determination of the local refractive index using Mach-Zehnder interferometry. Presented here are data for five protein and three virus systems. From data for these and other systems, optical monitoring experiments to measure local growth conditions and growth kinetics in liquid-liquid diffusion, batch, and vapor diffusion crystal growth experiments can be designed
Recommended from our members
Isolation, characterization, and preliminary X-ray diffraction data for a serine protease from Penicillium cyclopium.
The major extracellular protein of Penicillium cyclopium has been isolated from its culture media and purified by ammonium sulfate fractionation, gel, and ion-exchange chromatography. We show this secreted protein to be endopeptidase. The molecular weight is approximately 32,000, the pI is 5.0, and the pH optimum using a variety of protein and synthetic substrates is around 7.0. Inhibition studies show that the protease is not inhibited by pepstatin nor by p-chloromercuribenzoic acid, indicating, respectively, that it is not an aspartyl protease nor a thiol protease. Complete inhibition is observed, however, with phenylmethanesulfonyl fluoride. Three crystal forms suitable for high resolution x-ray diffraction studies have been obtained from this purified protease with reflections being observed to well beyond 3.0 A resolution. One form having a needle morphology is of the orthorhombic crystal class and has space group P2(1)2(1)2(1). The unit cell dimensions are a = 41.9 A, b = 43.2 A, and c = 111.5 A with 1 molecule of the protease occurring in the asymmetric unit. The second form grown at pH values less than 6.0 has a plate morphology, is of orthorhombic space group P2(1)2(1)2(1), and has unit cell dimensions a = 59.12 A, b = 62.33 A, and c = 70.62 A. The third form is polyhedral in habit, is also of space group P2(1)2(1)2(1), and appears when the pH of the mother liquor is greater than 7.0. The cell dimensions of this crystal form are a = 57.07 A, b = 58.82 A, c = 70.79 A, and again there is 1 molecule/asymmetric unit. Three-dimensional structural analysis by x-ray diffraction is now underway. All crystal forms are somewhat denser than the norm having mass to volume ratios of 1.58, 2.00, and 1.85 A3/dalton, respectively
Recommended from our members
Isolation, characterization, and preliminary X-ray diffraction data for a serine protease from Penicillium cyclopium.
The major extracellular protein of Penicillium cyclopium has been isolated from its culture media and purified by ammonium sulfate fractionation, gel, and ion-exchange chromatography. We show this secreted protein to be endopeptidase. The molecular weight is approximately 32,000, the pI is 5.0, and the pH optimum using a variety of protein and synthetic substrates is around 7.0. Inhibition studies show that the protease is not inhibited by pepstatin nor by p-chloromercuribenzoic acid, indicating, respectively, that it is not an aspartyl protease nor a thiol protease. Complete inhibition is observed, however, with phenylmethanesulfonyl fluoride. Three crystal forms suitable for high resolution x-ray diffraction studies have been obtained from this purified protease with reflections being observed to well beyond 3.0 A resolution. One form having a needle morphology is of the orthorhombic crystal class and has space group P2(1)2(1)2(1). The unit cell dimensions are a = 41.9 A, b = 43.2 A, and c = 111.5 A with 1 molecule of the protease occurring in the asymmetric unit. The second form grown at pH values less than 6.0 has a plate morphology, is of orthorhombic space group P2(1)2(1)2(1), and has unit cell dimensions a = 59.12 A, b = 62.33 A, and c = 70.62 A. The third form is polyhedral in habit, is also of space group P2(1)2(1)2(1), and appears when the pH of the mother liquor is greater than 7.0. The cell dimensions of this crystal form are a = 57.07 A, b = 58.82 A, c = 70.79 A, and again there is 1 molecule/asymmetric unit. Three-dimensional structural analysis by x-ray diffraction is now underway. All crystal forms are somewhat denser than the norm having mass to volume ratios of 1.58, 2.00, and 1.85 A3/dalton, respectively
Recommended from our members
Determination of local refractive index for protein and virus crystals in solution by Mach-Zehnder interferometry.
To establish the importance of, and quantitatively evaluate, the macromolecular concentration gradients in the neighborhood of growing protein, virus, and nucleic acid crystals, a convenient, accurate, and nonintrusive method has been devised. This approach should prove particularly relevant in the rigorous comparison of crystals grown in a conventional laboratory setting with those grown in a microgravity environment. The method is based on precise determination of the local refractive index using Mach-Zehnder interferometry. Presented here are data for five protein and three virus systems. From data for these and other systems, optical monitoring experiments to measure local growth conditions and growth kinetics in liquid-liquid diffusion, batch, and vapor diffusion crystal growth experiments can be designed
Recommended from our members
Crystallization and preliminary X-ray analysis of the vitamin D-binding protein from human serum
The vitamin D-binding protein from human serum has been crystallized from polyethylene glycol as a complex with 25-hydroxyvitamin D and examined by X-ray diffraction photography. The space group of the crystals is C2 with a = 203.0 A ̊, b = 75.8 A ̊, c = 90.9 A ̊ and β = 109.5°. There are two molecules of 56,000 dalton in the asymmetric unit. The crystals diffract to about 3.0 Å resolution but the patterns exhibit a substantial level of diffuse scatter. © 1985.
Protein and virus crystal growth on international microgravity laboratory-2.
Two T = 1 and one T = 3 plant viruses, along with a protein, were crystallized in microgravity during the International Microgravity Laboratory-2 (IML-2) mission in July of 1994. The method used was liquid-liquid diffusion in the European Space Agency's Advanced Protein Crystallization Facility (APCF). Distinctive alterations in the habits of Turnip Yellow Mosaic Virus (TYMV) crystals and hexagonal canavalin crystals were observed. Crystals of cubic Satellite Tobacco Mosaic Virus (STMV) more than 30 times the volume of crystals grown in the laboratory were produced in microgravity. X-ray diffraction analysis demonstrated that both crystal forms of canavalin and the cubic STMV crystals diffracted to significantly higher resolution and had superior diffraction properties as judged by relative Wilson plots. It is postulated that the establishment of quasi-stable depletion zones around crystals growing in microgravity are responsible for self-regulated and more ordered growth