title: A novel method for simultaneous production of two ribosome-inactivating proteins, α-MMC and MAP30, from Momordica charantia L.
abstract: Alpha-momorcharin (α-MMC) and momordica anti-HIV protein (MAP30) from Momordica charantia L. have been confirmed to possess anti-tumor and anti-virus activities. Traditional purification methods of these two ribosome-inactivating proteins (RIPs) were separate which was time consuming and cost effective as well as low efficient. In order to obtain sufficient samples for researches, a strategy combining ion-exchange and gel filtration chromatography was developed and optimized in this study. Using this novel purification method, averagely 1162 mg of α-MMC and 535 mg of MAP30 were obtained from 400 g of Momordica charantia L seeds. The homogeneities of them were assessed by electrophoresis analysis. Determination of molecular weights of α-MMC and MAP30 were 28.585 kDa and 29.094 kDa by MALDI-TOF/TOF and pI were 9.02 and 9.12, respectively. The single glycoproteins were identified by Periodate-Schiff's base (PAS) and the saccharide content was tested to be 1.25% and 1.1% by anthrone-sulfuric acid method. Biological activities were evidenced by their ability to inhibit proliferation of lung adenocarcinoma A549 cell and to convert supercoiled plasmid pUC18 into relaxed forms. Finally, we also found that both two RIPs exhibited no superoxide dismutase (SOD) activity.
authors.affiliation: School of Medical Laboratory Science, Chengdu Medical College, Chengdu, China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Meng
authors.affiliation: Department of Histology and Embryology and Neurobiology, Development and Regeneration Key Laboratory of Sichuan Province, Chengdu Medical College, Chengdu, China.
authors.forename: Sen
authors.initials: S
authors.lastname: Lin
authors.affiliation: School of Medical Laboratory Science, Chengdu Medical College, Chengdu, China.
authors.forename: Shuangfeng
authors.initials: S
authors.lastname: Liu
authors.affiliation: Key Laboratory of Bio-resources and Eco-environment Ministry of Education/Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, College of Life Science, Sichuan University, Chengdu, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Fan
authors.affiliation: Key Laboratory of Bio-resources and Eco-environment Ministry of Education/Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, College of Life Science, Sichuan University, Chengdu, China.
authors.forename: Gangrui
authors.initials: G
authors.lastname: Li
authors.affiliation: Key Laboratory of Bio-resources and Eco-environment Ministry of Education/Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, College of Life Science, Sichuan University, Chengdu, China.
authors.forename: Yanfa
authors.initials: Y
authors.lastname: Meng
mesh_terms: D019380:Anti-HIV Agents / Q000302:isolation & purification*; D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D029777:Momordica charantia / Q000737:chemistry*; D054788:Ribosome Inactivating Proteins / Q000302:isolation & purification*; D054790:Ribosome Inactivating Proteins, Type 2 / Q000302:isolation & purification*; D012639:Seeds / Q000737:chemistry*; D013482:Superoxide Dismutase / Q000737:chemistry
chemical_list: D019380:Anti-HIV Agents; D000972:Antineoplastic Agents, Phytogenic; C066347:MAP30 protein, Momordica charantia; D054790:Ribosome Inactivating Proteins, Type 2; C039582:MMC protein, Momordica charantia; D013482:Superoxide Dismutase; D054788:Ribosome Inactivating Proteins