UCLA researchers in the Department of Pharmacology have developed a novel human exon library for mRNA display detection of protein-protein interactions.
BACKGROUND:
Proteins control all biological functions within a cell. The majority of proteins require interaction with other proteins as partners, to achieve their proper function in cellular machineries. Protein-protein interactions (PPIs) are vital for the pathology of viruses and other infectious agents, and therefore represent attractive therapeutic targets for anti-viral drugs. The most common strategy for identification of these PPIs is affinity-purification mass spectrometry (AP-MS). Although recent technological advances in AP-MS have led to increased accuracy of peptide detection, current strategies for PPI identification remain restricted by detection limits that often omit low abundance proteins. There is a need for novel methodologies capable of low abundance protein binder detection, for the mapping of PPIs to specific domains with increased specificity and sensitivity.
INNOVATION:
UCLA researchers have created an exon mRNA display method that enables the detection of low abundance PPIs with localization of the interaction sites to specific regions, with high specificity and sensitivity. The method is complementary to existing methodologies for PPI discovery, which will allow the simultaneous detection of low abundance interactors and interaction domain mapping. Moreover, the use of high-throughput DNA sequencing as the readout for this method permits sensitive quantification of interactions. This technique may be used for the identification of novel therapeutic targets at protein interaction interfaces.
POTENTIAL APPLICATIONS:
• Detection of low abundance protein-protein interactions
• Identification of novel therapeutic targets at viral-host interaction sites
ADVANTAGES:
• Detection of low abundance binders/interactors simultaneously with large-scale interaction domain mapping
• Mapping of PPIs to specific domains with increased specificity and sensitivity
• Sensitive quantification of interactions
DEVELOPMENT-TO-DATE:
This method has been applied to Influenza A Virus NS1, which validated previously described protein-protein interactions and identified novel cellular binders/interactors that are of low abundance in the cell.