title: Characterization of biochemical properties of feline foamy virus integrase.
abstract: In order to study biochemical properties, the integrase (IN) protein of feline foamy virus (FFV) was over-expressed from Escherichia coli, purified by two-step chromatography; Talon column and heparin column, and characterized in biochemical aspects. For the three enzymatic reactions of the 3' -processing, strand transfer, and disintegration activities, Mn2+ ion was essentially required as a cofactor. Interestingly, Co2+ and Zn2+ ions were found to act as effective cofactors, while other transition elements such as Ni2+, Cu2+, La3+, Y3+, Cd2+, Li1+, Ba2+, Sr2+, V3+, and so on were not. Regarding to the substrate specificity, FFV IN has low substrate specificities as it cleaved in a significant level prototype foamy virus (PFV) U5 LTR substrate as well as FFV U5 LTR substrate, while PFV IN did not. Finally, the 3' -processing activity was observed in the high concentrations of several solvents such as CHAPS, Glycerol, Tween 20 and Triton X-100, which are generally used for dissolution of chemicals in inhibitor-screening. Therefore, as it is the first report showing biochemical properties, FFV IN is proposed to have low specificities on the use of cofactor and substrate for enzymatic reaction when it is compared with other retroviral INs.
authors.affiliation: Department of Biotechnology, Chung-Ang University, Ansung 456-756, Korea.
authors.forename: Donghyun
authors.initials: D
authors.lastname: Lee
authors.forename: Usok
authors.initials: U
authors.lastname: Hyun
authors.forename: Ji-Ye
authors.initials: JY
authors.lastname: Kim
authors.forename: Cha-Gyun
authors.initials: CG
authors.lastname: Shin
mesh_terms: D004795:Enzyme Stability; D019426:Integrases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016092:Spumavirus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D014764:Viral Proteins; D019426:Integrases