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Glycosylation at Asn(211) Regulates the Activation State of the Discoidin Domain Receptor 1 (DDR1)

Discoidin domain receptor 1 (DDR1) belongs to a unique family of receptor tyrosine kinases that signal in response to collagens. DDR1 undergoes autophosphorylation in response to collagen binding with a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family....

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Autores principales: Fu, Hsueh-Liang, Valiathan, Rajeshwari R., Payne, Leo, Kumarasiri, Malika, Mahasenan, Kiran V., Mobashery, Shahriar, Huang, Paul, Fridman, Rafael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3979393/
https://www.ncbi.nlm.nih.gov/pubmed/24509848
http://dx.doi.org/10.1074/jbc.M113.541102
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author Fu, Hsueh-Liang
Valiathan, Rajeshwari R.
Payne, Leo
Kumarasiri, Malika
Mahasenan, Kiran V.
Mobashery, Shahriar
Huang, Paul
Fridman, Rafael
author_facet Fu, Hsueh-Liang
Valiathan, Rajeshwari R.
Payne, Leo
Kumarasiri, Malika
Mahasenan, Kiran V.
Mobashery, Shahriar
Huang, Paul
Fridman, Rafael
author_sort Fu, Hsueh-Liang
collection PubMed
description Discoidin domain receptor 1 (DDR1) belongs to a unique family of receptor tyrosine kinases that signal in response to collagens. DDR1 undergoes autophosphorylation in response to collagen binding with a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family. DDR1 dimerization precedes receptor activation suggesting a structural inhibitory mechanism to prevent unwarranted phosphorylation. However, the mechanism(s) that maintains the autoinhibitory state of the DDR1 dimers is unknown. Here, we report that N-glycosylation at the Asn(211) residue plays a unique role in the control of DDR1 dimerization and autophosphorylation. Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. Mass spectrometry revealed that the N211Q mutant undergoes phosphorylation at Tyr(484), Tyr(520), Tyr(792), and Tyr(797). The N211Q traffics to the cell surface, and its ectodomain displays collagen I binding with an affinity similar to that of the wild-type DDR1 ectodomain. However, unlike the wild-type receptor, the N211Q mutant exhibits enhanced receptor dimerization and sustained activation upon ligand withdrawal. Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. The presence of glycan moieties at that site may help to lock the collagen-binding domain in the inactive state and prevent unwarranted signaling by receptor dimers. These studies provide a novel insight into the structural mechanisms that regulate DDR activation.
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spelling pubmed-39793932014-04-09 Glycosylation at Asn(211) Regulates the Activation State of the Discoidin Domain Receptor 1 (DDR1) Fu, Hsueh-Liang Valiathan, Rajeshwari R. Payne, Leo Kumarasiri, Malika Mahasenan, Kiran V. Mobashery, Shahriar Huang, Paul Fridman, Rafael J Biol Chem Glycobiology and Extracellular Matrices Discoidin domain receptor 1 (DDR1) belongs to a unique family of receptor tyrosine kinases that signal in response to collagens. DDR1 undergoes autophosphorylation in response to collagen binding with a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family. DDR1 dimerization precedes receptor activation suggesting a structural inhibitory mechanism to prevent unwarranted phosphorylation. However, the mechanism(s) that maintains the autoinhibitory state of the DDR1 dimers is unknown. Here, we report that N-glycosylation at the Asn(211) residue plays a unique role in the control of DDR1 dimerization and autophosphorylation. Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. Mass spectrometry revealed that the N211Q mutant undergoes phosphorylation at Tyr(484), Tyr(520), Tyr(792), and Tyr(797). The N211Q traffics to the cell surface, and its ectodomain displays collagen I binding with an affinity similar to that of the wild-type DDR1 ectodomain. However, unlike the wild-type receptor, the N211Q mutant exhibits enhanced receptor dimerization and sustained activation upon ligand withdrawal. Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. The presence of glycan moieties at that site may help to lock the collagen-binding domain in the inactive state and prevent unwarranted signaling by receptor dimers. These studies provide a novel insight into the structural mechanisms that regulate DDR activation. American Society for Biochemistry and Molecular Biology 2014-03-28 2014-02-07 /pmc/articles/PMC3979393/ /pubmed/24509848 http://dx.doi.org/10.1074/jbc.M113.541102 Text en © 2014 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version full access. Creative Commons Attribution Unported License (http://creativecommons.org/licenses/by/3.0/) applies to Author Choice Articles
spellingShingle Glycobiology and Extracellular Matrices
Fu, Hsueh-Liang
Valiathan, Rajeshwari R.
Payne, Leo
Kumarasiri, Malika
Mahasenan, Kiran V.
Mobashery, Shahriar
Huang, Paul
Fridman, Rafael
Glycosylation at Asn(211) Regulates the Activation State of the Discoidin Domain Receptor 1 (DDR1)
title Glycosylation at Asn(211) Regulates the Activation State of the Discoidin Domain Receptor 1 (DDR1)
title_full Glycosylation at Asn(211) Regulates the Activation State of the Discoidin Domain Receptor 1 (DDR1)
title_fullStr Glycosylation at Asn(211) Regulates the Activation State of the Discoidin Domain Receptor 1 (DDR1)
title_full_unstemmed Glycosylation at Asn(211) Regulates the Activation State of the Discoidin Domain Receptor 1 (DDR1)
title_short Glycosylation at Asn(211) Regulates the Activation State of the Discoidin Domain Receptor 1 (DDR1)
title_sort glycosylation at asn(211) regulates the activation state of the discoidin domain receptor 1 (ddr1)
topic Glycobiology and Extracellular Matrices
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3979393/
https://www.ncbi.nlm.nih.gov/pubmed/24509848
http://dx.doi.org/10.1074/jbc.M113.541102
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