Home > Publications database > Expression, purification and membrane reconstitution of a CD4 fragment comprising the transmembrane and cyctoplasmic domains of the receptor > print

001     60123
005     20200402210439.0
024 7 _ |2 pmid
|a pmid:17613246
024 7 _ |2 DOI
|a 10.1016/j.pep.2007.05.007
024 7 _ |2 WOS
|a WOS:000249631800023
037 _ _ |a PreJuSER-60123
041 _ _ |a eng
082 _ _ |a 570
084 _ _ |2 WoS
|a Biochemical Research Methods
084 _ _ |2 WoS
|a Biochemistry & Molecular Biology
084 _ _ |2 WoS
|a Biotechnology & Applied Microbiology
100 1 _ |a Wittlich, M.
|b 0
|u FZJ
|0 P:(DE-Juel1)VDB28257
245 _ _ |a Expression, purification and membrane reconstitution of a CD4 fragment comprising the transmembrane and cyctoplasmic domains of the receptor
260 _ _ |a Orlando, Fla.
|b Academic Press
|c 2007
300 _ _ |a 198 - 207
336 7 _ |a Journal Article
|0 PUB:(DE-HGF)16
|2 PUB:(DE-HGF)
336 7 _ |a Output Types/Journal article
|2 DataCite
336 7 _ |a Journal Article
|0 0
|2 EndNote
336 7 _ |a ARTICLE
|2 BibTeX
336 7 _ |a JOURNAL_ARTICLE
|2 ORCID
336 7 _ |a article
|2 DRIVER
440 _ 0 |a Protein Expression and Purification
|x 1046-5928
|0 18221
|v 55
500 _ _ |a Record converted from VDB: 12.11.2012
520 _ _ |a The 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.
536 _ _ |a Funktion und Dysfunktion des Nervensystems
|c P33
|2 G:(DE-HGF)
|0 G:(DE-Juel1)FUEK409
|x 0
588 _ _ |a Dataset connected to Web of Science, Pubmed
650 _ 2 |2 MeSH
|a Amino Acid Sequence
650 _ 2 |2 MeSH
|a Antigens, CD4: biosynthesis
650 _ 2 |2 MeSH
|a Antigens, CD4: genetics
650 _ 2 |2 MeSH
|a Antigens, CD4: isolation & purification
650 _ 2 |2 MeSH
|a Cell Membrane: chemistry
650 _ 2 |2 MeSH
|a Escherichia coli: genetics
650 _ 2 |2 MeSH
|a Genes, Synthetic
650 _ 2 |2 MeSH
|a Humans
650 _ 2 |2 MeSH
|a Liposomes: chemistry
650 _ 2 |2 MeSH
|a Molecular Sequence Data
650 _ 2 |2 MeSH
|a Nuclear Magnetic Resonance, Biomolecular
650 _ 2 |2 MeSH
|a Protein Structure, Tertiary
650 _ 2 |2 MeSH
|a Recombinant Proteins: biosynthesis
650 _ 2 |2 MeSH
|a Recombinant Proteins: chemistry
650 _ 2 |2 MeSH
|a Recombinant Proteins: isolation & purification
650 _ 2 |2 MeSH
|a Viral Regulatory and Accessory Proteins: chemistry
650 _ 7 |0 0
|2 NLM Chemicals
|a Antigens, CD4
650 _ 7 |0 0
|2 NLM Chemicals
|a Liposomes
650 _ 7 |0 0
|2 NLM Chemicals
|a Recombinant Proteins
650 _ 7 |0 0
|2 NLM Chemicals
|a Viral Regulatory and Accessory Proteins
650 _ 7 |a J
|2 WoSType
653 2 0 |2 Author
|a CD4
653 2 0 |2 Author
|a membrane protein
653 2 0 |2 Author
|a HIV-1
653 2 0 |2 Author
|a NMR spectroscopy
653 2 0 |2 Author
|a ubiquitin-fusion system
700 1 _ |a Wiesehan, K.
|b 1
|u FZJ
|0 P:(DE-Juel1)VDB15437
700 1 _ |a Koenig, B. W.
|b 2
|u FZJ
|0 P:(DE-Juel1)132009
700 1 _ |a Willbold, D.
|b 3
|u FZJ
|0 P:(DE-Juel1)132029
773 _ _ |a 10.1016/j.pep.2007.05.007
|g Vol. 55, p. 198 - 207
|p 198 - 207
|q 55<198 - 207
|0 PERI:(DE-600)1471688-4
|t Protein expression and purification
|v 55
|y 2007
|x 1046-5928
856 7 _ |u http://dx.doi.org/10.1016/j.pep.2007.05.007
909 C O |o oai:juser.fz-juelich.de:60123
|p VDB
913 1 _ |k P33
|v Funktion und Dysfunktion des Nervensystems
|l Funktion und Dysfunktion des Nervensystems
|b Gesundheit
|0 G:(DE-Juel1)FUEK409
|x 0
914 1 _ |y 2007
915 _ _ |0 StatID:(DE-HGF)0010
|a JCR/ISI refereed
920 1 _ |k INB-2
|l Molekulare Biophysik
|d 31.12.2008
|g INB
|0 I:(DE-Juel1)VDB805
|x 0
920 1 _ |k JARA-SIM
|l Jülich-Aachen Research Alliance - Simulation Sciences
|g JARA
|0 I:(DE-Juel1)VDB1045
|x 1
970 _ _ |a VDB:(DE-Juel1)94323
980 _ _ |a VDB
980 _ _ |a ConvertedRecord
980 _ _ |a journal
980 _ _ |a I:(DE-Juel1)ISB-2-20090406
980 _ _ |a I:(DE-Juel1)VDB1045
980 _ _ |a UNRESTRICTED
980 _ _ |a I:(DE-Juel1)ICS-6-20110106
981 _ _ |a I:(DE-Juel1)IBI-7-20200312
981 _ _ |a I:(DE-Juel1)ISB-2-20090406
981 _ _ |a I:(DE-Juel1)VDB1045
981 _ _ |a I:(DE-Juel1)ICS-6-20110106

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