Wong Jack H, Ng T B
Department of Biochemistry, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China.
Peptides. 2003 Jul;24(7):963-8. doi: 10.1016/s0196-9781(03)00192-x.
From the seeds of the Yunnan bean, we purified an antifungal peptide using affinity chromatography on Affi-gel blue gel, FPLC-ion exchange chromatography on Mono S, and FPLC-gel filtration on Superdex 75. The antifungal peptide was adsorbed on Affi-gel blue gel at pH 7.8 and Mono S at pH 4.5. It exhibited a molecular mass of 6.5 kDa in both gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its N-terminal sequence closely resembled defensin-related peptides. The peptide exerted antifungal activity toward the fungal species Fusarium oxysporum and Mycosphaerella arachidicola, with an IC50 of 2 microM for the former fungus and 10 microM for the latter. It manifested a weaker mitogenic activity toward murine splenocytes than Concanavalin A. It also displayed antiproliferative activity on a murine leukemia (L1210), a hepatoma (HepG2), and a murine leukemia (M1) cell line. It inhibited human immunodeficiency virus-1 reverse transcriptase with an IC50 of 200 microM.
我们从云南豆种子中，通过在Affi - 凝胶蓝胶上进行亲和层析、在Mono S上进行快速蛋白质液相色谱离子交换层析以及在Superdex 75上进行快速蛋白质液相色谱凝胶过滤，纯化出一种抗真菌肽。该抗真菌肽在pH 7.8时吸附于Affi - 凝胶蓝胶，在pH 4.5时吸附于Mono S。在凝胶过滤和十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳中，其分子量均为6.5 kDa。它的N端序列与防御素相关肽非常相似。该肽对尖孢镰刀菌和落花生球腔菌具有抗真菌活性，对前者的IC50为2 microM，对后者为10 microM。与伴刀豆球蛋白A相比，它对小鼠脾细胞的促有丝分裂活性较弱。它还对小鼠白血病（L1210）、肝癌（HepG2）和小鼠白血病（M1）细胞系表现出抗增殖活性。它以200 microM的IC50抑制人类免疫缺陷病毒1逆转录酶。
