PATHOGEN RECEPTOR DISCOVERY WITH A MICROFLUIDIC HUMAN MEMBRANE PROTEIN ARRAY

The discovery of how a pathogen invades a cell requires one to determine which host cell receptors are exploited. This is a challenging problem in protein-protein interaction not least because the receptor is invariably a membrane protein, which represent an Achilles heel in proteomics. We have developed a generalized platform for high throughput expression and interaction studies of membrane proteins by creating a microfluidic-based comprehensive human membrane protein array (MPA). The MPA offers a powerful alternative to conventional proteomics by enabling the simultaneous study of 2,100 membrane proteins. We characterized direct interactions of a whole virus (SV40) with candidate host receptors expressed on MPA, and discovered candidate receptors of which eighteen were validated, thus demonstrating that this first of its kind application is an important tool for cellular receptor discovery and the understanding of pathogen tropism. Considering the tremendous difficulty in discovering pathogen receptors, this in vitro high throughput approach is extremely important for receptor discovery and understanding pathogen tropism, with relevance to emerging human diseases.