Structure-based mutagenesis of the substrate-recognition domain of Nrdp1/FLRF identifies the binding site for the receptor tyrosine kinase ErbB3

Samuel Bouyain, Daniel J. Leahy

Research output: Contribution to journalArticle

Abstract

The E3 ubiquitin ligase neuregulin receptor degrading protein 1 (Nrdp1) mediates the ligand-independent degradation of the epidermal growth factor receptor family member ErbB3/HER3. By regulating cellular levels of ErbB3, Nrdp1 influences ErbB3-mediated signaling, which is essential for normal vertebrate development. Nrdp1 belongs to the tripartite or RBCC (RING, B-box, coiled-coil) family of ubiquitin ligases in which the RING domain is responsible for ubiquitin ligation and a variable C-terminal region mediates substrate recognition. We report here the 1.95 Å crystal structure of the C-terminal domain of Nrdp1 and show that this domain is sufficient to mediate ErbB3 binding. Furthermore, we have used site-directed mutagenesis to map regions of the Nrdp1 surface that are important for interacting with ErbB3 and mediating its degradation in transfected cells. The ErbB3-binding site localizes to a region of Nrdp1 that is conserved from invertebrates to vertebrates, in contrast to ErbB3, which is only found in vertebrates. This observation suggests that Nrdp1 uses a common binding site to recognize its targets in different species. Published by Cold Spring Harbor Laboratory Press.

Original languageEnglish (US)
Pages (from-to)654-661
Number of pages8
JournalProtein Science
Volume16
Issue number4
DOIs
StatePublished - Apr 2007

Fingerprint

Neuregulins
Mutagenesis
Receptor Protein-Tyrosine Kinases
Binding Sites
Substrates
Proteins
Vertebrates
Ubiquitin
Degradation
Ubiquitin-Protein Ligases
Invertebrates
Ligases
Ports and harbors
Site-Directed Mutagenesis
Epidermal Growth Factor Receptor
Ligation
Membrane Proteins
Crystal structure
Ligands

Keywords

  • Crystal structure
  • E3 ubiquitin ligase
  • ErbB3
  • Protein degradation
  • Receptor tyrosine kinase

ASJC Scopus subject areas

  • Biochemistry

Cite this

Structure-based mutagenesis of the substrate-recognition domain of Nrdp1/FLRF identifies the binding site for the receptor tyrosine kinase ErbB3. / Bouyain, Samuel; Leahy, Daniel J.

In: Protein Science, Vol. 16, No. 4, 04.2007, p. 654-661.

Research output: Contribution to journalArticle

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