Linked Life Data

8003258

Source:http://linkedlifedata.com/resource/pubmed/id/8003258

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
1994-7-20
pubmed:abstractText
Homodimers of immunoglobulin VL domains are minimal models of antibodies in that they display an ensemble of six hypervariable loops. Bence Jones protein REI is a mixture of a complete kappa light chain and the corresponding variable domain (REIV). The known three-dimensional structure of the REIV dimer (Epp et al., 1975, Biochemistry 14, 4943-4952) provides a basis for studying dimer stabilization by protein engineering. Mutant REIV-L94H was constructed and shown to have an equilibrium constant of dimerization about one order of magnitude higher than wildtype REIV. By fusing REIV and variants to the aminoterminal part of the Vibrio cholerae ToxR regulator protein (Miller et al., 1987, Cell 48, 271-279), a transcriptional signal in E. coli can be derived from REIV homodimer formation constant. The system senses dimerization of the immunoglobulin part of the fusion protein, located in the periplasmatic space, and transduces the signal as transcriptional activation to a ctx::lacZ gene construct integrated into the E. coli chromosome. There is positive correlation between the propensities of homodimer formation and the rate of transcriptional initiation at the ctx promoter. Since beta-galactosidase levels can easily be measured colorimetrically in crude cell lysates of a large number of clones using an ELISA reader, this procedure constitutes all elements required for a genetic screen in E. coli for immunoglobulin variants with altered association constants.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0177-3593
pubmed:author
pubmed:issnType
Print
pubmed:volume
375
pubmed:geneSymbol
rei, toxR
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
61-70
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:8003258-Bacterial Proteins, pubmed-meshheading:8003258-Base Sequence, pubmed-meshheading:8003258-Bence Jones Protein, pubmed-meshheading:8003258-Chromatography, Gel, pubmed-meshheading:8003258-DNA-Binding Proteins, pubmed-meshheading:8003258-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:8003258-Escherichia coli, pubmed-meshheading:8003258-Genetic Vectors, pubmed-meshheading:8003258-Models, Genetic, pubmed-meshheading:8003258-Models, Molecular, pubmed-meshheading:8003258-Molecular Sequence Data, pubmed-meshheading:8003258-Polymers, pubmed-meshheading:8003258-Promoter Regions, Genetic, pubmed-meshheading:8003258-Recombinant Fusion Proteins, pubmed-meshheading:8003258-Transcription, Genetic, pubmed-meshheading:8003258-Transcription Factors, pubmed-meshheading:8003258-Vibrio cholerae
pubmed:year
1994
pubmed:articleTitle
Dimerization of Bence Jones proteins: linking the rate of transcription from an Escherichia coli promoter to the association constant of REIV.
pubmed:affiliation
Institut für Molekulare Genetik, Georg-August-Universität Göttingen, Germany.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't