Proteolytic fragments of anti-HIV and anti-tumor proteins MAP30 and GAP31 are biologically active

被引:48
作者
Huang, PL
Sun, YT
Chen, HC
Kung, HF
Huang, PL
Lee-Huang, S [1 ]
机构
[1] NYU, Sch Med, Dept Biochem, New York, NY 10016 USA
[2] Amer Biosci, New York, NY 10021 USA
[3] NICHHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA
[4] Univ Hong Kong, Inst Mol Biol, Hong Kong, Peoples R China
[5] Massachusetts Gen Hosp, Dept Med, Boston, MA 02114 USA
[6] Harvard Univ, Sch Med, Boston, MA 02114 USA
关键词
D O I
10.1006/bbrc.1999.1213
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We analyzed the structural and functional organization of anti-HIV and anti-tumor proteins MAP30 and GAP31 by limited proteolysis with endopeptidases Lys-C and Glu-C (V8), MAP30 and GAP31 are resistant to proteolytic digestion under conditions of as much as 5% (w/w) proteases. In the presence of 10% (w/w) protease, the central regions of the proteins are still. resistant to proteolysis, whereas the N- and C-termini are accessible. Peptide fragments were purified by FPLC on Superdex 75 columns, characterized by gel electrophoresis, identified by amino acid sequencing, and analyzed for anti-HIV, anti-tumor, and other biochemical activities. We report here that limited proteolysis yields biologically active fragments of both MAP30 and GAP31. These fragments are active against HIV-1 and tumor cells with EC(50)s in the sub-nanomolar ranges, 0.2-0.4 nM. At the dose levels used in the assays, little cytotoxicity to normal cells was observed. In addition, these fragments remain fully active in HIV-integrase inhibition and HIV-LTR topological inactivation, but not ribosome inactivation. These results demonstrate that the antiviral and anti-tumor activities of MAP30 and GAP31 are independent of ribosome inactivation activity. In addition, we demonstrate that portions of the N- and C-termini are not essential for antiviral and anti-tumor activities, but do appear to be required for ribosome inactivation. These results may provide novel strategies for rational design and targeted development of mimetic antiviral and anti-tumor therapeutics, (C) 1999 Academic Press.
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收藏
页码:615 / 623
页数:9
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