Mono-PEGylation of Alpha-MMC and MAP30 from Momordica charantia L.: Production, Identification and Anti-Tumor Activity.
PEGylation is a well-established and effective strategy to decrease immunogenicity, which can increase the stability and in vivo half-life time. However, the generation of multi-site modified products is inevitable due to the lysine chemistry, which will bring difficulties in subsequent research, such as purification and quantification. Site-specific modification by mPEG-succinimidyl carbonate (mPEG-SC) is a widely used method forN-terminal conjugation. In this study, we used it for site-directed modification on two ribosome-inactivating proteins (RIPs), alpha-momorcharin (α-MMC) and momordica anti-HIV protein (MAP30), fromMomordica charantiaL. According to the optimization of previous modification conditions, we compared Macro-Cap SP with SP-Sepharose FF chromatography for separating the final mPEGylated RIPs. Two kinds of methods both can obtain homogenous mPEGylated RIPs which were identified by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing electrophoresis (IEF), and matrix-assisted laser desorption ionization-time of flight/time of flight (MALDI-TOF/TOF) analysis. We also used iodine staining method to detect the amount of unmodified PEG. Furthermore, the inhibition activity of both mPEGylated and non-PEGylated RIPs against human lung adenocarcinoma epithelial A549 cells was detected. All of the results suggested that the mPEGylated α-MMC/MAP30 might be potentially developed as new anti-tumor drugs.
School of Medical Laboratory Science, Chengdu Medical College, Chengdu 610500, Sichuan, China. sy83016603@163.com.|The First Affiliated Hospital of Chengdu Medical College, Chengdu 610000, Sichuan, China. sy83016603@163.com.
Yun
Y
Sun
School of Medical Laboratory Science, Chengdu Medical College, Chengdu 610500, Sichuan, China. sunfenghui_83@hotmail.com.
Fenghui
F
Sun
School of Medical Laboratory Science, Chengdu Medical College, Chengdu 610500, Sichuan, China. jlli999@foxmail.com.
Jianlong
J
Li
School of Medical Laboratory Science, Chengdu Medical College, Chengdu 610500, Sichuan, China. wml8198@sina.com.
Minlu
M
Wu
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 610064, Sichuan, China. fxscu1990@163.com.
Xiang
X
Fan
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 610064, Sichuan, China. yfmeng0902@scu.edu.cn.
Yanfa
Y
Meng
School of Medical Laboratory Science, Chengdu Medical College, Chengdu 610500, Sichuan, China. myaoworks@foxmail.com.
Yao
Y
Meng
D000230:Adenocarcinoma / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D000972:Antineoplastic Agents, Phytogenic / Q000737:chemistry / Q000494:pharmacology; D045744:Cell Line, Tumor; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D029777:Momordica charantia / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry*; D054790:Ribosome Inactivating Proteins, Type 2 / Q000737:chemistry / Q000494:pharmacology
D000972:Antineoplastic Agents, Phytogenic; C066347:MAP30 protein, Momordica charantia; D054790:Ribosome Inactivating Proteins, Type 2; D011092:Polyethylene Glycols; C000595209:polyethylene glycol 1000
alpha-momorcharin (α-MMC); anti-tumor; momordica anti-HIV protein (MAP30); protein PEGylation; ribosome-inactivating protein