Core Services

 

 

 

 

 

 

 

 

 

 

 

 

 

Nucleic Acids Programmable Protein Array Core (NAPPA)

(Funding for this core ended March 1, 2011)

Former Core leader: Grant McFadden

University of Florida

 

The goal of the Core was to assist the deciphering of the molecular mechanisms of host responses to select pathogens using proteomics to uncover novel protein-protein interactions (PPIs). This project was originally pursued in collaboration with the Harvard Proteomics Facility, which was developing the NAPPA technology to create human and select bacterial pathogen protein microarrays. However, our pilot screenings yielded far too many false-positive PPI hits, using the NAPPA technology as adapted for multi-well format, and we switched to an alternative technology, called Alphascreen, that utilizes the same plasmid library that was created for NAPPA. In contrast to NAPPA, which utilizes in vitro expression of proteins, the Alphascreen method utilizes proteins expressed in vivo within mammalian cells. Our collaborators included several SERCEB investigators, including Drs. Mark Dennison and Ralph Baric (human coronaviruses), Dr. Margo Brinton (West Nile Virus [WNV]), Dr. Aravinda deSilva (Dengue viruses), Dr. Mark Heise (Chikungunya [CHIKV], Sindbis, Ross River, and Venezuelan Equine Encephalitis [VEE] viruses), and Dr. David Weiss (F. tularensis). We screened these pathogen proteins for novel PPIs against host protein targets that are elements of the human inflammasome complexes and the RIG-I pathway. Specifically, we: