A novel ribonuclease with HIV-1 reverse transcriptase inhibitory activity purified from the fungus Ramaria formosa

被引:12
作者
Zhang, Rui [1 ,2 ]
Tian, Guoting [3 ]
Zhao, Yongchang [3 ]
Zhao, Liyan [4 ]
Wang, Hexiang [1 ,2 ]
Gong, Zhiyuan [5 ]
Ng, Tzi Bun [6 ]
机构
[1] China Agr Univ, State Key Lab Agrobiotechnol, Beijing 100193, Peoples R China
[2] China Agr Univ, Dept Microbiol, Beijing 100193, Peoples R China
[3] Yunnan Acad Agr Sci, Inst Biotechnol & Germplasm Resource, Kunming, Peoples R China
[4] Nanjing Agr Univ, Coll Food Sci & Technol, Nanjing, Jiangsu, Peoples R China
[5] Shandong Acad Agr Sci, Inst Agr Resources & Environm, Jinan, Shandong, Peoples R China
[6] Chinese Univ Hong Kong, Sch Biomed Sci, Fac Med, Shatin, Hong Kong, Peoples R China
关键词
Ribonuclease; HIV-1 reverse transcriptase; Ramaria formosa; FRESH FRUITING BODIES; HUMAN-IMMUNODEFICIENCY-VIRUS; UBIQUITIN-LIKE PEPTIDE; ANTIPROLIFERATIVE ACTIVITY; SEMINAL RIBONUCLEASE; PURIFICATION; PROTEIN; EXPRESSION; MUSHROOMS; ANTIFUNGAL;
D O I
10.1002/jobm.201300876
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A purification protocol that encompassed anion exchange chromatography (EC) on DEAE-cellulose, cation EC on CM-cellulose, anion EC on Q-Sepharose, and fast protein liquid chromatography-gel filtration of Superdex 75 was used to isolate a ribonuclease from dried fruiting bodies of Ramaria formosa. The ribonuclease was unadsorbed on CM-cellulose but adsorbed on both DEAE-cellulose and Q-Sepharose. It displayed a molecular mass of 29-kDa in both gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The ranking of its ribonucleolytic activity toward polyhomoribonucleotides was poly(U)>poly(C)>poly(G)>poly(A). It exhibited a pH optimum of pH 5 and a temperature optimum at 60 degrees C. The ribonuclease inhibited HIV-1 reverse transcriptase with an IC50 of 3 mu M.
引用
收藏
页码:269 / 275
页数:7
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