Lee-Huang S, Huang P L, Chen H C, Huang P L, Bourinbaiar A, Huang H I, Kung H F
Department of Biochemistry, New York University School of Medicine, New York 10016, USA.
Gene. 1995 Aug 19;161(2):151-6. doi: 10.1016/0378-1119(95)00186-a.
MAP30 is an anti-HIV plant protein that we have identified and purified to homogeneity from bitter melon (Momordica charantia). It is capable of acting against multiple stages of the viral life cycle, on acute infection as well as replication in chronically infected cells. In addition to antiviral action, MAP30 also possesses anti-tumor activity, topological inactivation of viral DNA, inhibition of viral integrase and cell-free ribosome-inactivation activities. We have cloned and expressed the MAP30 gene. The objective of this study is to characterize recombinant MAP30 (re-MAP30) and to determine its anti-HIV, anti-tumor and other activities. We report here that re-MAP30 inhibits HIV-1 and certain human tumors to the same extent as its native counterpart, natural MAP30 (nMAP30). The anti-HIV activity was measured by quantitative focal syncytium formation on CEM-ss cell monolayers, viral core protein p24 expression and viral-associated reverse transcriptase activity in HIV-1-infected H9 cells. The anti-tumor activity was measured by metabolic labeling of protein synthesis in tumor cells. In the dose range of the assay, re-MAP30 exhibits little toxicity to the uninfected viral target cells and other normal human cells. Identical to nMAP30, re-MAP30 is also active in topological inactivation of viral DNA, inhibition of viral DNA integration and cell-free ribosome inactivation. The cloning and expression of the gene encoding biologically active re-MAP30 provides an abundant source of homogeneous material for clinical investigations, as well as structure-function studies of this novel antiviral and anti-tumor agent.
MAP30是一种抗HIV植物蛋白，我们已从苦瓜（苦瓜属）中鉴定并纯化至同质。它能够作用于病毒生命周期的多个阶段，包括急性感染以及在慢性感染细胞中的复制。除了抗病毒作用外，MAP30还具有抗肿瘤活性、病毒DNA的拓扑失活、病毒整合酶的抑制以及无细胞核糖体失活活性。我们已经克隆并表达了MAP30基因。本研究的目的是表征重组MAP30（re-MAP30）并确定其抗HIV、抗肿瘤及其他活性。我们在此报告，re-MAP30抑制HIV-1和某些人类肿瘤的程度与天然对应物天然MAP30（nMAP30）相同。通过在CEM-ss细胞单层上进行定量灶性合胞体形成、HIV-1感染的H9细胞中病毒核心蛋白p24表达和病毒相关逆转录酶活性来测量抗HIV活性。通过肿瘤细胞中蛋白质合成的代谢标记来测量抗肿瘤活性。在测定的剂量范围内，re-MAP30对未感染的病毒靶细胞和其他正常人类细胞几乎没有毒性。与nMAP30相同，re-MAP30在病毒DNA的拓扑失活、病毒DNA整合的抑制和无细胞核糖体失活方面也具有活性。编码具有生物活性的re-MAP30的基因的克隆和表达为临床研究以及这种新型抗病毒和抗肿瘤药物的结构-功能研究提供了丰富的同质材料来源。
