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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
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pubmed:dateCreated |
1996-12-4
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pubmed:abstractText |
Recombinant Nef-protein of HIV-1 Bru derived from Escherichia coli revealed heparin-binding activity. This property was used to purify the Nef-protein by a one-step procedure, yielding about 90% homogenous Nef-protein as evaluated by silver staining. The Nef-protein was soluble without denaturing agents. Native folding of Nef was demonstrated with antibodies against conformational epitopes of Nef by a slot blot assay under native conditions. Despite its affinity to heparin and its nuclear localization in persistently HIV-1 infected glioblastoma cells (Kohleisen et al., 1992), Nef did not show DNA-binding properties by slot blot/hybridization assay and South/Western blot. In nucleotide-binding assays a strong autophosphorylation activity with [gamma-32P]ATP was observed. Nef-protein was not a substrate for ADP-ribosylation by bacterial toxins arguing against G-protein-like activities of Nef. Recombinant Nef did not interact with membranes as shown by the lack of increased fluorescence emission of Nef in the presence of liposomes. The recombinant Nef-protein obtained by one-step heparin-based purification shares immunological properties with native Nef and should prove useful for further studies of Nef function and immunogenicity.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Antibodies, Monoclonal,
http://linkedlifedata.com/resource/pubmed/chemical/DNA,
http://linkedlifedata.com/resource/pubmed/chemical/Gene Products, nef,
http://linkedlifedata.com/resource/pubmed/chemical/HIV Antibodies,
http://linkedlifedata.com/resource/pubmed/chemical/Heparin,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Fusion Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/nef Gene Products, Human...
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pubmed:status |
MEDLINE
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pubmed:month |
Jun
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pubmed:issn |
0166-0934
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
60
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
89-101
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pubmed:dateRevised |
2007-11-15
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pubmed:meshHeading |
pubmed-meshheading:8795010-Animals,
pubmed-meshheading:8795010-Antibodies, Monoclonal,
pubmed-meshheading:8795010-Cell Membrane,
pubmed-meshheading:8795010-DNA,
pubmed-meshheading:8795010-Gene Products, nef,
pubmed-meshheading:8795010-HIV Antibodies,
pubmed-meshheading:8795010-HIV-1,
pubmed-meshheading:8795010-Heparin,
pubmed-meshheading:8795010-Humans,
pubmed-meshheading:8795010-Mice,
pubmed-meshheading:8795010-Rabbits,
pubmed-meshheading:8795010-Recombinant Fusion Proteins,
pubmed-meshheading:8795010-Tumor Cells, Cultured,
pubmed-meshheading:8795010-nef Gene Products, Human Immunodeficiency Virus
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pubmed:year |
1996
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pubmed:articleTitle |
Heparin-binding capacity of the HIV-1 NEF-protein allows one-step purification and biochemical characterization.
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pubmed:affiliation |
GSF-Institut für Molekulare Virologie, Neuherberg, Oberschleissheim, FRG.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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