SMART-TMT

Since 2012, we have been profiling protein interactions in human cells via affinity-purification mass spectrometry and systematically analyzing interactions for all accessible human proteins at proteome scale. Leveraging the clones available in the human ORFeome (v. 8.1) developed by Marc Vidal and David Hill at the Dana Farber Cancer Center, we have been expressing C-terminally HA-FLAG-tagged versions of each human protein for immunopurification and LC-MS identification of binding partners. We have then been combining these interaction profiles by the thousands to create a series of models of the human interactome with steadily increasing scale. While the first, BioPlex 1.0 - HEK293T v1.0, included 24,000 interactions among 8,000 proteins, BioPlex 2.0 - HEK293T v2.0 expanded coverage to 57,000 interactions among 11,000 proteins. The most recent, BioPlex 3.0 - HEK293T v3.0 & HCT116 v1.0, includes nearly 120,000 interactions among nearly 15,000 proteins and is the most comprehensive..