1. Purification and characterization of a novel ubiquitin-like antitumour protein with hemagglutinating and deoxyribonuclease activities from the edible mushroom Ramaria botrytis
- Author
-
Fang Liu, Ke-Ren Zhang, Ya-Jie Han, Tzi Bun Ng, Rong Zhou, and Min-Hui Zhang
- Subjects
0301 basic medicine ,Antitumour protein ,030102 biochemistry & molecular biology ,biology ,Hemagglutination ,Ramaria botrytis ,Biophysics ,Deoxyribonuclease ,Apoptosis ,biology.organism_classification ,Applied Microbiology and Biotechnology ,Molecular biology ,Edible mushroom ,03 medical and health sciences ,030104 developmental biology ,Ubiquitin ,Biochemistry ,Annexin ,Sephadex ,biology.protein ,Original Article ,Ultracentrifuge ,Peptide sequence - 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.
- Published
- 2017