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• W2004221867 endingPage "829" @default.
• W2004221867 startingPage "819" @default.
• W2004221867 abstract "The diversity and domain structure of αβ T cell receptors (TCR) are similar to immunoglobulins based on sequence homologies, but the three-dimensional structure of the αβ-heterodimer has not been solved. To begin structure/function studies, we have compared the properties of a soluble single-chain VαVβ TCR (scTCR) expressed in three E. coli systems. The Vα and Vβ regions were expressed with pelB or ompA signal sequences or as a thioredoxin fusion protein. The scTCRs were detected only in the insoluble fraction of the cells and could be solubilized in guanidine and renatured to obtain properly folded scTCR from each system. Only a small fraction (1–5%) of the ompA and pelB scTCRs folded properly. In contrast, the thioredoxin fusion protein exhibited high total yields and a solubility that was ten times higher than the other scTCRs. The thioredoxin fusion protein also bound specifically to the peptide/MHC ligand with a KD of ∼0.7 μM, as shown by a competitive inhibition assay with Fab fragments that recognize the MHC complex. Furthermore, estimates from saturation binding with antibodies that react with the native TCR indicated that up to 80% of the thioredoxin fusion protein was in the properly folded from. The improved yield, solubility, and binding activity of the thioredoxin-scTCR should make it useful for various structure/function studies." @default.
• W2004221867 created "2016-06-24" @default.
• W2004221867 creator A5040656659 @default.
• W2004221867 creator A5088135588 @default.
• W2004221867 creator A5090805347 @default.
• W2004221867 date "1996-06-01" @default.
• W2004221867 modified "2023-10-17" @default.
• W2004221867 title "Binding properties and solubility of single-chain T cell receptors expressed in E. coli" @default.
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• W2004221867 doi "https://doi.org/10.1016/0161-5890(96)00038-7" @default.
• W2004221867 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/8811077" @default.
• W2004221867 hasPublicationYear "1996" @default.
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