Defining the binding interface of Amyloid Precursor Protein (APP) and Contactin3 (CNTN3) by site-directed mutagenesis

Xi Peng, John Williams, Philip M. Smallwood, Jeremy Nathans

Research output: Contribution to journalArticle

Abstract

The Amyloid Precursor Protein (APP) and Contactin (CNTN) families of cell-surface proteins have been intensively studied in the context of neural development and neuropsychiat-ric diseases. Earlier studies demonstrated both genetic and biochemical interactions between the extracellular domains of APP and CNTN3, but their precise binding interfaces were not defined. In the present study, we have used binding assays between APP-alkaline phosphatase (AP) fusion proteins and CNTN-Fc fusion proteins, together with alanine substitution mutagenesis, to show that: (i) the second Fibronectin domain (Fn(2)) in CNTN3 mediates APP binding; (ii) the copper binding domain (CuBD) in APP mediates CNTN3 binding; and (iii) the most important amino acids for APP-CNTN3 binding reside on one face of CNTN3-Fn(2) and on one face of APP-CuBD. These experiments define the regions of direct contact that mediate the binding interaction between APP and CNTN3.

Original languageEnglish (US)
Article numbere0219384
JournalPloS one
Volume14
Issue number7
DOIs
StatePublished - Jan 1 2019

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Mutagenesis
Amyloid beta-Protein Precursor
site-directed mutagenesis
amyloid
Site-Directed Mutagenesis
Contactins
Amyloid
Protein Binding
proteins
protein binding
Fusion reactions
Phosphoprotein Phosphatases
Fibronectins
Alanine
Alkaline Phosphatase
Copper
Molecular Biology
Membrane Proteins
Proteins
neurodevelopment

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Defining the binding interface of Amyloid Precursor Protein (APP) and Contactin3 (CNTN3) by site-directed mutagenesis. / Peng, Xi; Williams, John; Smallwood, Philip M.; Nathans, Jeremy.

In: PloS one, Vol. 14, No. 7, e0219384, 01.01.2019.

Research output: Contribution to journalArticle

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