Collect. Czech. Chem. Commun.
1996, 61, 1254-1260
https://doi.org/10.1135/cccc19961254
Effect of Solvent Composition on Solubility and Activity of Bovine Leukemia Virus Proteinase
Olga Hrušková-Heidingsfeldováa, Martin Piskáčekb, Romana Cubínkováa, Aleš Zábranskýa and Iva Pichováa
a Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, 166 10 Prague 6, Czech Republic
b Department of Biochemistry and Microbiology, Prague Institute of Chemical Technology, 166 28 Prague 6, Czech Republic
Abstract
The solubility and activity of bovine leukemia virus proteinase (BLV PR) has been examined as a function of solvent composition in order to find a buffer system which would enable to prepare samples concentrated sufficiently for crystallization attempts and other studies. Low solubility and frequent precipitation was one of the major difficulties in the course of experiments carried out with BLV PR in several laboratories. We found that buffers composed of polyvalent ions, and containing glycerol, ethylene glycol or dimethyl sulfoxide, enabled to concentrate the enzyme up to 2 mg/ml. Presence of 4-20% of DMSO, isopropyl alcohol or ethylene glycol results in activation of BLV PR, while glycerol does not significantly affect the BLV PR activity. Zinc or magnesium ions substituting for sodium ions in commonly used buffers caused decrease both in solubility and activity of BLV PR.
Keywords: BLV proteinase; Specific activity; Solubility; Solvent composition.
