Purification and characterization of a novel antifungal protein with antiproliferation and anti-HIV-1 reverse transcriptase activities from Peganum harmala seeds  

作　　者：Xiaojin Ma Dongliang Liu Haishu Tang Yan Wang Ting Wu Yang Li Jie Yang Jianhua Yang Surong Sun Fuchun Zhang 

机构地区：[1]Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China [2]Department of Pediatrics, Texas Children's Cancer Center, Dan L. Duncan Cancer Center, Baylor College of Medicine, Houston, TX, USA

出　　处：《Acta Biochimica et Biophysica Sinica》2013年第2期87-94,共8页生物化学与生物物理学报（英文版）

摘　　要：A novel antifungal protein, designated as PHP, was isolated from the seeds of Peganum harmala, by cationic exchange chromatography on Resource S column and gel filtration on Sephadex 75 10/300 GL column. PHP was found to form a homodimer of about 16 kDa. Isoelectric focusing polyacrylamide gel electrophoresis analysis showed that the isoelectric point of PHP was - 8.4. The N-terminal 20-amino acid sequence of PHP, ITCPQVTQSLAPCVPYLISG, resembles the non-specific lipid transfer proteins in certain plants. PHP exhibited lipid-binding activity. Furthermore, PHP exerted antifungal activity against Alternaria alternate, Penicillium degitatum, Rhizopus stuolonifer, and Magnaporthe grisea, and its antifungal activity was stable in the temperature range 4-60℃, and in the pH range 4-10.It inhibited the mycelial growth in A. alternate, P. degitatum, R. stuolonifer, and M. grisea with 50% inhibitory concentration （IC50） of 1.5, 37.5, 8.44, and 12.19 μM, respectively.PHP was also able to inhibit the proliferation of esophagus carcinoma （Eca-109）, cervical carcinoma （HeLa）, gastric carcinoma （MGC-7）, and melanoma （B16） cells with IC50 of 0.7, 2.74, 3.13, and 1.47μM, respectively. Moreover, PHP significantly inhibited HIV-1 reverse transcriptase （RT） with an IC50 of 1.26 μM. It did not have hemagglutinating and antibacterial activities. In conclusion, a novel antifungal protein with antiproliferation and anti-HIV-1 RT activities was obtained from P. harmala seeds.A novel antifungal protein, designated as PHP, was isolated from the seeds of Peganum harmala, by cationic exchange chromatography on Resource S column and gel filtration on Sephadex 75 10/300 GL column. PHP was found to form a homodimer of about 16 kDa. Isoelectric focusing polyacrylamide gel electrophoresis analysis showed that the isoelectric point of PHP was - 8.4. The N-terminal 20-amino acid sequence of PHP, ITCPQVTQSLAPCVPYLISG, resembles the non-specific lipid transfer proteins in certain plants. PHP exhibited lipid-binding activity. Furthermore, PHP exerted antifungal activity against Alternaria alternate, Penicillium degitatum, Rhizopus stuolonifer, and Magnaporthe grisea, and its antifungal activity was stable in the temperature range 4-60℃, and in the pH range 4-10.It inhibited the mycelial growth in A. alternate, P. degitatum, R. stuolonifer, and M. grisea with 50% inhibitory concentration （IC50） of 1.5, 37.5, 8.44, and 12.19 μM, respectively.PHP was also able to inhibit the proliferation of esophagus carcinoma （Eca-109）, cervical carcinoma （HeLa）, gastric carcinoma （MGC-7）, and melanoma （B16） cells with IC50 of 0.7, 2.74, 3.13, and 1.47μM, respectively. Moreover, PHP significantly inhibited HIV-1 reverse transcriptase （RT） with an IC50 of 1.26 μM. It did not have hemagglutinating and antibacterial activities. In conclusion, a novel antifungal protein with antiproliferation and anti-HIV-1 RT activities was obtained from P. harmala seeds.

关 键 词：Peganum harmala lipid transfer proteins antifungal activity ANTITUMOR anti-HIV-1 reversetranscriptase activity 

分 类 号：Q78[生物学—分子生物学] Q966