A Protein-Fragment Complementation Assay (PCA) is a protein-protein interaction assay in which a bait protein is expressed as fusion to one of the either N- or C- terminal peptide fragments of a reporter protein and prey protein is expressed as fusion to the complementary N- or C- terminal fragment of the same reporter protein. Interaction of bait and prey proteins bring together complementary fragments, which can then fold into an active reporter, e.g. the split-ubiquitin assay.

Publication

Monitoring protein-protein interactions in mammalian cells by trans-SUMOylation.

Srivastav RK, Schwede S, Klaus M, Schwermann J, Gaestel M, Niedenthal R

Protein-protein interactions are essential for almost all cellular processes, hence understanding these processes mainly depends on the identification and characterization of the relevant protein-protein interactions. In the present paper, we introduce the concept of TRS (trans-SUMOylation), a new method developed to identify and verify protein-protein interactions in mammalian cells in vivo. TRS utilizes Ubc9-fusion proteins that trans-SUMOylate co-expressed interacting proteins. ... [more]

Biochem. J. Sep. 15, 2011; 438(3);495-503 [Pubmed: 21675959]

Throughput

High Throughput

Additional Notes

interaction detected by trans-sumoylation
trans-sumoylation: bait protein is fused to Ubc9 and hit protein is sumoylated if that bait and hit protein interact

Related interactions

Interaction	Experimental Evidence Code	Dataset	Throughput	Score	Curated By	Notes
MAP2K2 MAPK1	Biochemical Activity Biochemical Activity An interaction is inferred from the biochemical effect of one protein upon another, for example, GTP-GDP exchange activity or phosphorylation of a substrate by a kinase. The bait protein executes the activity on the substrate hit protein. A Modification value is recorded for interactions of this type with the possible values Phosphorylation, Ubiquitination, Sumoylation, Dephosphorylation, Methylation, Prenylation, Acetylation, Deubiquitination, Proteolytic Processing, Glucosylation, Nedd(Rub1)ylation, Deacetylation, No Modification, Demethylation.	Ho DT (2003)	Low	-	BioGRID	857936
MAP2K2 MAPK1	Reconstituted Complex Reconstituted Complex An interaction is inferred between proteins in vitro. This can include proteins in recombinant form or proteins isolated directly from cells with recombinant or purified bait. For example, GST pull-down assays where a GST-tagged protein is first isolated and then used to fish interactors from cell lysates are considered reconstituted complexes (e.g. PUBMED: 14657240, Fig. 4A or PUBMED: 14761940, Fig. 5). This can also include gel-shifts, surface plasmon resonance, isothermal titration calorimetry (ITC) and bio-layer interferometry (BLI) experiments. The bait-hit directionality may not be clear for 2 interacting proteins. In these cases the directionality is up to the discretion of the curator.	Ho DT (2003)	Low	-	BioGRID	-
MAP2K2 MAPK1	Two-hybrid Two-hybrid Bait protein expressed as a DNA binding domain (DBD) fusion and prey expressed as a transcriptional activation domain (TAD) fusion and interaction measured by reporter gene activation.	Robinson FL (2002)	Low	-	BioGRID	-

Curated By

BioGRID

Interaction Summary

MAPK1

Gene Ontology Biological Process

Gene Ontology Molecular Function

RNA polymerase II carboxy-terminal domain kinase activity [ISS]phosphatase binding [IPI]protein binding [IPI]protein serine/threonine kinase activity [IDA]

Gene Ontology Cellular Component

MAP2K2