Zhao Yu, Jin Yang, Wang Jian-Hua, Wang Rui-Rui, Yang Liu-Meng, Lee Wen-Hui, Zheng Yong-Tang, Zhang Yun
Department of Animal Toxinology, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan 650223, China.
Toxicon. 2005 Nov;46(6):619-24. doi: 10.1016/j.toxicon.2005.06.022. Epub 2005 Sep 12.
A novel protein, named BAS-AH, was purified and characterized from the skin of the toad Bufo andrewsi. BAS-AH is a single chain protein and the apparent molecular weight is about 63 kDa as judged by SDS-PAGE. BAS-AH was determined to bind heme (0.89 mol heme/mol protein) as determined by pyridine haemochrome analysis. Fifty percentage cytotoxic concentration (CC50) of BAS-AH on C8166 cells was 9.5 microM. However, at concentrations that showed little effect on cell viability, BAS-AH displayed dose dependent inhibition on HIV-1 infection and replication. The antiviral selectivity indexes (CC50/EC50) were 14.4 and 11.4, respectively, corresponding to the measurements of syncytium formation and HIV-1 p24 antigen expression. BAS-AH also showed an inhibitory effect on the activity of recombinant HIV-1 reverse transcriptase (IC50 = 1.32 microM). The N-terminal sequence of BAS-AH was determined to be NAKXKADVIGKISILLGQDNLSNIVAAM, which exhibited little identity with other known anti-HIV-1 proteins. BAS-AH is devoid of antibacterial, proteolytic, trypsin inhibitory activity, l-amino acid oxidase activity and catalase activity.
从中华大蟾蜍皮肤中纯化并鉴定出一种名为BAS - AH的新型蛋白质。BAS - AH是一种单链蛋白质，通过SDS - PAGE判断其表观分子量约为63 kDa。通过吡啶血色原分析确定BAS - AH可结合血红素（0.89摩尔血红素/摩尔蛋白质）。BAS - AH对C8166细胞的半数细胞毒性浓度（CC50）为9.5 microM。然而，在对细胞活力影响较小的浓度下，BAS - AH对HIV - 1感染和复制表现出剂量依赖性抑制作用。抗病毒选择性指数（CC50/EC50）分别为14.4和11.4，对应于对合胞体形成和HIV - 1 p24抗原表达的测量。BAS - AH对重组HIV - 1逆转录酶的活性也有抑制作用（IC50 = 1.32 microM）。BAS - AH的N端序列确定为NAKXKADVIGKISILLGQDNLSNIVAAM，与其他已知的抗HIV - 1蛋白几乎没有同源性。BAS - AH没有抗菌、蛋白水解、胰蛋白酶抑制活性、L - 氨基酸氧化酶活性和过氧化氢酶活性。
