Dimerization of Soluble Disulfide Trap Single-Chain Major Histocompatibility Complex Class I Molecules Dependent on Peptide Binding Affinity

2011 ◽  
Vol 15 (3) ◽  
pp. 635-644
Author(s):  
Eleni Kotsiou ◽  
Joanna Brzostek ◽  
Izabela Lenart ◽  
Antony N. Antoniou ◽  
Julian Dyson ◽  
...  
Keyword(s):  
Major Histocompatibility Complex ◽  
Major Histocompatibility Complex Class ◽  
Binding Affinity ◽  
Peptide Binding ◽  
Class I ◽  
Complex Class ◽  
Major Histocompatibility ◽  
Single Chain ◽  
Histocompatibility Complex ◽  
Peptide Binding Affinity

scholarly journals Destabilizing single chain major histocompatibility complex class I protein for repurposed enterokinase proteolysis

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Jackwee Lim
Keyword(s):  
Major Histocompatibility Complex ◽  
Major Histocompatibility Complex Class ◽  
De Novo ◽  
Class I ◽  
Complex Class ◽  
Major Histocompatibility ◽  
Single Chain ◽  
Mhc I ◽  
Histocompatibility Complex ◽  
Protein Instability

Abstract The lack of a high throughput assay for screening stabilizing peptides prior to building a library of peptide-major histocompatibility complex class I (pMHC-I) molecules has motivated the continual use of in silico tools without biophysical characterization. Here, based on de novo protein fragmentation, the EASY MHC-I (EZ MHC-I) assay favors peptide antigen screening to an unheralded hands-on time of seconds per peptide due to the empty single chain MHC-I protein instability. Unlike tedious traditional labeling- and antibody-based MHC-I assays, repurposed enterokinase directly fragments the unstable single MHC-I chain protein unless rescued by a stabilizing peptide under luminal condition. Herein, the principle behind EZ MHC-I assay not only characterizes the overlooked stability as a known better indicator of immunogenicity than classical affinity but also the novel use of enterokinase from the duodenum to target destabilized MHC-I protein not bearing the standard Asp-Asp-Asp-Asp-Lys motif, which may protend to other protein instability-based assays.

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