title: Purification and characterization of a ribonuclease with antiproliferative activity from the mystical wild mushroom Tuber indicum.
abstract: An RNase with a molecular mass of 28 kDa and with high ribonucleolytic activity toward poly(A) was purified from the ascocarps of Tuber indicum. The purification procedure involved ion exchange chromatography on diethylaminoethyl cellulose, Q-Sepharose and Mono Q, and gel filtration by fast protein liquid chromatography on Superdex 75. The pH and temperature optima of the RNase were 7.2 and 50 °C, respectively. The ranking of its activity toward various polyhomoribonucleotides was poly(A)>poly(C)>poly(G) ≈ poly(U). All of the metal ions used in this study, except for the K(+) ions, curtailed the activity of the RNase. The RNase activity was reduced by ethylene diamine tetraacetic acid (EDTA), dithiothreitol (DTT), and sodium dodecyl sulfate (SDS) by 42.2%, 75.5%, and 96.6%, respectively. The RNase inhibited the proliferation of hepatoma (HepG2) and human breast cancer cell lines (MCF7), with half-maximal inhibitory concentrations (IC50 ) of 12.6 and 16.6 μM, respectively.
authors.affiliation: State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Xiao
authors.forename: ShanShan
authors.initials: S
authors.lastname: Feng
authors.forename: HeXiang
authors.initials: H
authors.lastname: Wang
authors.forename: ZhiYuan
authors.initials: Z
authors.lastname: Gong
authors.forename: TziBun
authors.initials: T
authors.lastname: Ng
mesh_terms: D000363:Agaricales / Q000201:enzymology*; D001203:Ascomycota / Q000201:enzymology*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004791:Enzyme Inhibitors / Q000378:metabolism; D004795:Enzyme Stability; D006801:Humans; D006863:Hydrogen-Ion Concentration; D020128:Inhibitory Concentration 50; D008670:Metals / Q000378:metabolism; D008970:Molecular Weight; D011061:Poly A / Q000378:metabolism*; D012260:Ribonucleases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D004791:Enzyme Inhibitors; D008670:Metals; D011061:Poly A; D012260:Ribonucleases
keywords: Antiproliferative; Purification; Ribonuclease; Truffle; Tuber indicum