Periplasmic expression of soluble single chain T cell receptors is rescued by the chaperone FkpA

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Periplasmic expression of soluble single chain T cell receptors is rescued by the chaperone FkpA

BACKGROUND Efficient expression systems exist for antibody (Ab) molecules, which allow for characterization of large numbers of individual Ab variants. In contrast, such expression systems have been lacking for soluble T cell receptors (TCRs). Attempts to generate bacterial systems have generally resulted in low yields and material which is prone to aggregation and proteolysis. Here we present ...

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Periplasmic chaperone FkpA is essential for imported colicin M toxicity

Chaperones facilitate correct folding of newly synthesized proteins. We show here that the periplasmic FkpA chaperone is required for killing Escherichia coli by colicin M entering cells from the outside. Highly active colicin M preparations were inactive against fkpA mutant cells; 10(4)-fold dilutions killed fkpA(+) cells. Three previously isolated spontaneous mutants tolerant to colicin M car...

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The periplasmic Escherichia coli peptidylprolyl cis,trans-isomerase FkpA. II. Isomerase-independent chaperone activity in vitro.

We recently identified FkpA by selecting for the increased yield of antibody single-chain Fv (scFv) fragments in phage display, even of those not containing cis-prolines. We have now investigated the properties of FkpA in vitro. The peptidylprolyl cis-trans-isomerase activity of FkpA was found to be among the highest of any such enzyme with a protein substrate, yet FkpA is not able to enhance t...

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Functional three-domain single-chain T-cell receptors.

T-cell receptors (TCRs) are membrane anchored heterodimers structurally related to antibody molecules. Single-chain antibodies can be engineered by linking the two variable domains, which fold properly by themselves. However, proper assembly of the variable domains of a human TCR (V alpha and V beta) that recognize the HLA-DR2b/myelin basic protein-(85-99) peptide complex was critically depende...

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Soluble T cell receptors: novel immunotherapies.

T cell receptors are antigen-specific proteins that have evolved to recognize peptide antigens presented by human leukocyte antigen molecules on most cell types. Like antibodies, T cell receptors are produced with huge diversity but, unlike antibodies, T cell receptors are not secreted and do not undergo somatic mutations that increase their affinities for antigen. Recently, however, methods ha...

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ژورنال

عنوان ژورنال: BMC Biotechnology

سال: 2010

ISSN: 1472-6750

DOI: 10.1186/1472-6750-10-8