New Partners Identified by Mass Spectrometry Assay Reveal Functions of NCAM2 in Neural Cytoskeleton Organization

Int J Mol Sci. 2021 Jul 9;22(14):7404. doi: 10.3390/ijms22147404.

Abstract

Neuronal cell adhesion molecule 2 (NCAM2) is a membrane protein with an important role in the morphological development of neurons. In the cortex and the hippocampus, NCAM2 is essential for proper neuronal differentiation, dendritic and axonal outgrowth and synapse formation. However, little is known about NCAM2 functional mechanisms and its interactive partners during brain development. Here we used mass spectrometry to study the molecular interactome of NCAM2 in the second postnatal week of the mouse cerebral cortex. We found that NCAM2 interacts with >100 proteins involved in numerous processes, including neuronal morphogenesis and synaptogenesis. We validated the most relevant interactors, including Neurofilaments (NEFs), Microtubule-associated protein 2 (MAP2), Calcium/calmodulin kinase II alpha (CaMKIIα), Actin and Nogo. An in silico analysis of the cytosolic tail of the NCAM2.1 isoform revealed specific phosphorylation site motifs with a putative affinity for some of these interactors. Our results expand the knowledge of NCAM2 interactome and confirm the key role of NCAM2 in cytoskeleton organization, neuronal morphogenesis and synaptogenesis. These findings are of interest in explaining the phenotypes observed in different pathologies with alterations in the NCAM2 gene.

Keywords: CaMKIIα; MAP2; NCAM2; Neurofilaments; Nogo; cytoskeleton; mass spectrometry; neuronal morphogenesis.

MeSH terms

  • Actins / metabolism
  • Animals
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2 / metabolism
  • Cerebral Cortex / growth & development
  • Cerebral Cortex / metabolism*
  • Computational Biology
  • Cytoplasm / genetics
  • Cytoplasm / metabolism
  • Cytoskeleton / metabolism*
  • Databases, Chemical
  • Gene Ontology
  • In Vitro Techniques
  • Intermediate Filaments / metabolism
  • Mass Spectrometry*
  • Mice
  • Microtubule-Associated Proteins / metabolism
  • Neural Cell Adhesion Molecules / metabolism*
  • Neurogenesis* / genetics
  • Neurons / metabolism*
  • Nogo Proteins
  • Phosphorylation
  • Protein Domains
  • Protein Interaction Maps
  • Proteome / genetics
  • Proteome / metabolism
  • Transcriptome / genetics

Substances

  • Actins
  • Microtubule-Associated Proteins
  • Mtap2 protein, mouse
  • Ncam2 protein, mouse
  • Neural Cell Adhesion Molecules
  • Nogo Proteins
  • Proteome
  • Rtn4 protein, mouse
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2