title: Purification and characterization of a novel ubiquitin-like antitumour protein with hemagglutinating and deoxyribonuclease activities from the edible mushroom Ramaria botrytis.
abstract: A novel ubiquitin-like antitumour protein (RBUP) was isolated from fruiting bodies of the edible mushroom Ramaria botrytis. The protein was isolated with a purification protocol involving ion exchange chromatography on DEAE-Sepharose fast flow and gel filtration on Sephadex G-75. SDS-PAGE, Native-PAGE and ultracentrifugation analysis disclosed that RBUP was a monomeric protein with a molecular weight of 18.5 kDa. ESI-MS/MS demonstrated that it shared 69% amino acid sequence similarity with Coprinellus congregates ubiquitin (gi|136667). The protein exhibiting strong anticancer activity towards A549 cells. Analysis by employing AO/EB staining and Annexin V-FITC/PI detection indicated that the cytotoxic effect of RBUP was mediated through induction of apoptosis. Furthermore, RBUP displayed hemagglutinating and deoxyribonuclease activities. A temperature of 40 °C and pH of 7.0 were required for optimal DNase activity. Therefore, it was estimated that RBUP exerted its antitumour effect by inducing apoptosis, and its hemagglutinating and DNase activities were also thought to participate in this effect. These results demonstrated that RBUP was a multifunctional protein with potential medicinal applications.
authors.affiliation: The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Nankai University, Tianjin, 300071, China.
authors.forename: Rong
authors.initials: R
authors.lastname: Zhou
authors.affiliation: The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Nankai University, Tianjin, 300071, China.
authors.forename: Ya-Jie
authors.initials: YJ
authors.lastname: Han
authors.affiliation: The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Nankai University, Tianjin, 300071, China.
authors.forename: Min-Hui
authors.initials: MH
authors.lastname: Zhang
authors.affiliation: The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Nankai University, Tianjin, 300071, China.
authors.forename: Ke-Ren
authors.initials: KR
authors.lastname: Zhang
authors.affiliation: School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China. b021770@mailserv.cuhk.edu.hk.
authors.forename: Tzi Bun
authors.initials: TB
authors.lastname: Ng
authors.affiliation: The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Nankai University, Tianjin, 300071, China. liufang312@nankai.edu.cn.
authors.forename: Fang
authors.initials: F
authors.lastname: Liu
keywords: Antitumour protein; Apoptosis; Deoxyribonuclease; Hemagglutination; Ramaria botrytis