Spatial localisation of Discoidin Domain Receptor 2 (DDR2) signalling is dependent on its collagen binding and kinase activity.
Abstract
Discoidin Domain Receptor 2 (DDR2) is a collagen-binding receptor tyrosine kinase that initiates delayed and sustained tyrosine phosphorylation signalling. To understand the molecular basis of this unique phosphorylation profile, here we utilise fluorescence microscopy to map the spatiotemporal localisation of DDR2 and tyrosine phosphorylated proteins upon stimulation with collagen. We show that cellular phosphorylated proteins are localised to the interface where DDR2 is in contact with collagen and not in the early endosomes or lysosomes. We find that DDR2 localisation is independent of integrin activation and the key DDR2 signalling effector SHC1. Structure-function analysis reveals that DDR2 mutants defective for collagen binding or kinase activity are unable to localise to the cell surface, demonstrating for the first time that both collagen binding and kinase functions are required for spatial localisation of DDR2. This study provides new insights into the underlying structural features that control DDR2 activation in space and time.
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Subject
Cell Membrane
Humans
Collagen
Tyrosine
Integrins
Recombinant Proteins
Microscopy, Fluorescence
Amino Acid Substitution
Mutagenesis, Site-Directed
Signal Transduction
Protein Binding
Phosphorylation
Mutant Proteins
HEK293 Cells
Src Homology 2 Domain-Containing, Transforming Protein 1
Discoidin Domain Receptor 2
Research team
Protein Networks
Molecular and Systems Oncology
Language
eng
Date accepted
2018-04-24
License start date
2018-06
Citation
Biochemical and biophysical research communications, 2018, 501 (1), pp. 124 - 130
Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE