Bimolecular complementation affinity purification (BiCAP) reveals dimer-specific protein interactions for ERBB2 dimers

The dynamic assembly of multiprotein complexes is a central mechanism of many cell signaling pathways. This process is key to maintaining the spatiotemporal specificity required for an accurate, yet adaptive, response to rapidly changing cellular conditions. We describe a technique for the specific...

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Vydané v:Science signaling Ročník 9; číslo 436; s. ra69
Hlavní autori: Croucher, David R, Iconomou, Mary, Hastings, Jordan F, Kennedy, Sean P, Han, Jeremy Z R, Shearer, Robert F, McKenna, Jessie, Wan, Adrian, Lau, Joseph, Aparicio, Samuel, Saunders, Darren N
Médium: Journal Article
Jazyk:English
Vydavateľské údaje: United States 12.07.2016
Predmet:
Breast Neoplasms - metabolism
Breast Neoplasms - pathology
Female
HEK293 Cells
Humans
MAP Kinase Signaling System
MCF-7 Cells
Protein Multimerization
Receptor, Epidermal Growth Factor - isolation & purification
Receptor, Epidermal Growth Factor - metabolism
Receptor, ErbB-2 - isolation & purification
Receptor, ErbB-2 - metabolism
Receptor, ErbB-3 - isolation & purification
Receptor, ErbB-3 - metabolism
ISSN:1937-9145, 1937-9145
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Shrnutí:The dynamic assembly of multiprotein complexes is a central mechanism of many cell signaling pathways. This process is key to maintaining the spatiotemporal specificity required for an accurate, yet adaptive, response to rapidly changing cellular conditions. We describe a technique for the specific isolation and downstream proteomic characterization of any two interacting proteins, to the exclusion of their individual moieties and competing binding partners. We termed the approach bimolecular complementation affinity purification (BiCAP) because it combines the use of conformation-specific nanobodies with a protein-fragment complementation assay with affinity purification. Using BiCAP, we characterized the specific interactome of the epidermal growth factor receptor (EGFR) family member ERBB2 when in the form of a homodimer or when in the form of a heterodimer with either EGFR or ERBB3. We identified dimer-specific interaction patterns for key adaptor proteins and identified a number of previously unknown interacting partners. Functional analysis for one of these newly identified partners revealed a noncanonical mechanism of extracellular signal-regulated kinase (ERK) activation that is specific to the ERBB2:ERBB3 heterodimer and acts through the adaptor protein FAM59A in breast cancer cells.
Bibliografia:ObjectType-Article-1
SourceType-Scholarly Journals-1
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content type line 23
ISSN:1937-9145
1937-9145
DOI:10.1126/scisignal.aaf0793