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000060123 0247_ $$2pmid$$apmid:17613246
000060123 0247_ $$2DOI$$a10.1016/j.pep.2007.05.007
000060123 0247_ $$2WOS$$aWOS:000249631800023
000060123 037__ $$aPreJuSER-60123
000060123 041__ $$aeng
000060123 082__ $$a570
000060123 084__ $$2WoS$$aBiochemical Research Methods
000060123 084__ $$2WoS$$aBiochemistry & Molecular Biology
000060123 084__ $$2WoS$$aBiotechnology & Applied Microbiology
000060123 1001_ $$0P:(DE-Juel1)VDB28257$$aWittlich, M.$$b0$$uFZJ
000060123 245__ $$aExpression, purification and membrane reconstitution of a CD4 fragment comprising the transmembrane and cyctoplasmic domains of the receptor
000060123 260__ $$aOrlando, Fla.$$bAcademic Press$$c2007
000060123 300__ $$a198 - 207
000060123 3367_ $$0PUB:(DE-HGF)16$$2PUB:(DE-HGF)$$aJournal Article
000060123 3367_ $$2DataCite$$aOutput Types/Journal article
000060123 3367_ $$00$$2EndNote$$aJournal Article
000060123 3367_ $$2BibTeX$$aARTICLE
000060123 3367_ $$2ORCID$$aJOURNAL_ARTICLE
000060123 3367_ $$2DRIVER$$aarticle
000060123 440_0 $$018221$$aProtein Expression and Purification$$v55$$x1046-5928
000060123 500__ $$aRecord converted from VDB: 12.11.2012
000060123 520__ $$aThe transmembrane glycoprotein CD4 plays a prominent role in the adaptive immune response. CD4 is displayed primarily on the surface of T helper cells, but also on subsets of memory and regulatory T lymphocytes, macrophages, and dendritic cells. Binding of the lymphocyte specific tyrosine kinase p56(lck) to the cytoplasmic domain of CD4 is crucial for antigen receptor-mediated signal transduction. The human immunodeficiency virus (HIV) utilizes CD4 as the main receptor for T cell invasion. The virus has developed multiple strategies for down-regulation of CD4 in infected cells. Physical interactions of viral proteins VpU and Nef with the cytoplasmic tail of CD4 initiate a cascade of events leading to degradation of CD4. Here we report heterologous expression and purification of a CD4 fragment comprising the transmembrane and cytoplasmic domains of human CD4. A synthetic gene encoding CD4 amino acid residues 372-433 and a protease cleavage site was cloned into the pTKK19xb/ub plasmid. The CD4 fragment was expressed in Escherichia coli C43(DE3) cells as a ubiquitin fusion with an N-terminal His-tag, isolated, released by PreScission proteolytic cleavage, and purified to homogeneity. Incorporation of the recombinant CD4 fragment in lipid membranes and physical interaction with the cytoplasmic domain of VpU was demonstrated by centrifugation assays followed by reversed phase chromatographic analysis of the composition of the proteoliposomes. A high resolution NMR spectrum of uniformly (15)N-labeled CD4 peptide in membrane simulating micelles proves the possibility of solution NMR studies of this CD4 fragment and of its molecular complexes.
000060123 536__ $$0G:(DE-Juel1)FUEK409$$2G:(DE-HGF)$$aFunktion und Dysfunktion des Nervensystems$$cP33$$x0
000060123 588__ $$aDataset connected to Web of Science, Pubmed
000060123 650_2 $$2MeSH$$aAmino Acid Sequence
000060123 650_2 $$2MeSH$$aAntigens, CD4: biosynthesis
000060123 650_2 $$2MeSH$$aAntigens, CD4: genetics
000060123 650_2 $$2MeSH$$aAntigens, CD4: isolation & purification
000060123 650_2 $$2MeSH$$aCell Membrane: chemistry
000060123 650_2 $$2MeSH$$aEscherichia coli: genetics
000060123 650_2 $$2MeSH$$aGenes, Synthetic
000060123 650_2 $$2MeSH$$aHumans
000060123 650_2 $$2MeSH$$aLiposomes: chemistry
000060123 650_2 $$2MeSH$$aMolecular Sequence Data
000060123 650_2 $$2MeSH$$aNuclear Magnetic Resonance, Biomolecular
000060123 650_2 $$2MeSH$$aProtein Structure, Tertiary
000060123 650_2 $$2MeSH$$aRecombinant Proteins: biosynthesis
000060123 650_2 $$2MeSH$$aRecombinant Proteins: chemistry
000060123 650_2 $$2MeSH$$aRecombinant Proteins: isolation & purification
000060123 650_2 $$2MeSH$$aViral Regulatory and Accessory Proteins: chemistry
000060123 650_7 $$00$$2NLM Chemicals$$aAntigens, CD4
000060123 650_7 $$00$$2NLM Chemicals$$aLiposomes
000060123 650_7 $$00$$2NLM Chemicals$$aRecombinant Proteins
000060123 650_7 $$00$$2NLM Chemicals$$aViral Regulatory and Accessory Proteins
000060123 650_7 $$2WoSType$$aJ
000060123 65320 $$2Author$$aCD4
000060123 65320 $$2Author$$amembrane protein
000060123 65320 $$2Author$$aHIV-1
000060123 65320 $$2Author$$aNMR spectroscopy
000060123 65320 $$2Author$$aubiquitin-fusion system
000060123 7001_ $$0P:(DE-Juel1)VDB15437$$aWiesehan, K.$$b1$$uFZJ
000060123 7001_ $$0P:(DE-Juel1)132009$$aKoenig, B. W.$$b2$$uFZJ
000060123 7001_ $$0P:(DE-Juel1)132029$$aWillbold, D.$$b3$$uFZJ
000060123 773__ $$0PERI:(DE-600)1471688-4$$a10.1016/j.pep.2007.05.007$$gVol. 55, p. 198 - 207$$p198 - 207$$q55<198 - 207$$tProtein expression and purification$$v55$$x1046-5928$$y2007
000060123 8567_ $$uhttp://dx.doi.org/10.1016/j.pep.2007.05.007
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000060123 9131_ $$0G:(DE-Juel1)FUEK409$$bGesundheit$$kP33$$lFunktion und Dysfunktion des Nervensystems$$vFunktion und Dysfunktion des Nervensystems$$x0
000060123 9141_ $$y2007
000060123 915__ $$0StatID:(DE-HGF)0010$$aJCR/ISI refereed
000060123 9201_ $$0I:(DE-Juel1)VDB805$$d31.12.2008$$gINB$$kINB-2$$lMolekulare Biophysik$$x0
000060123 9201_ $$0I:(DE-Juel1)VDB1045$$gJARA$$kJARA-SIM$$lJülich-Aachen Research Alliance - Simulation Sciences$$x1
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