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Interactions between semaphorins and plexin–neuropilin receptor complexes in the membranes of live cells

Signaling of semaphorin ligands via their plexin–neuropilin receptors is involved in tissue patterning in the developing embryo. These proteins play roles in cell migration and adhesion but are also important in disease etiology, including in cancer angiogenesis and metastasis. While some structures...

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Autores principales: Christie, Shaun M., Hao, Jing, Tracy, Erin, Buck, Matthias, Yu, Jennifer S., Smith, Adam W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8350011/
https://www.ncbi.nlm.nih.gov/pubmed/34270956
http://dx.doi.org/10.1016/j.jbc.2021.100965
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author Christie, Shaun M.
Hao, Jing
Tracy, Erin
Buck, Matthias
Yu, Jennifer S.
Smith, Adam W.
author_facet Christie, Shaun M.
Hao, Jing
Tracy, Erin
Buck, Matthias
Yu, Jennifer S.
Smith, Adam W.
author_sort Christie, Shaun M.
collection PubMed
description Signaling of semaphorin ligands via their plexin–neuropilin receptors is involved in tissue patterning in the developing embryo. These proteins play roles in cell migration and adhesion but are also important in disease etiology, including in cancer angiogenesis and metastasis. While some structures of the soluble domains of these receptors have been determined, the conformations of the full-length receptor complexes are just beginning to be elucidated, especially within the context of the plasma membrane. Pulsed-interleaved excitation fluorescence cross-correlation spectroscopy allows direct insight into the formation of protein–protein interactions in the membranes of live cells. Here, we investigated the homodimerization of neuropilin-1 (Nrp1), plexin A2, plexin A4, and plexin D1 using pulsed-interleaved excitation fluorescence cross-correlation spectroscopy. Consistent with previous studies, we found that Nrp1, plexin A2, and plexin A4 are present as dimers in the absence of exogenous ligand. Plexin D1, on the other hand, was monomeric under similar conditions, which had not been previously reported. We also found that plexin A2 and A4 assemble into a heteromeric complex. Stimulation with semaphorin 3A or semaphorin 3C neither disrupts nor enhances the dimerization of the receptors when expressed alone, suggesting that activation involves a conformational change rather than a shift in the monomer–dimer equilibrium. However, upon stimulation with semaphorin 3C, plexin D1 and Nrp1 form a heteromeric complex. This analysis of interactions provides a complementary approach to the existing structural and biochemical data that will aid in the development of new therapeutic strategies to target these receptors in cancer.
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spelling pubmed-83500112021-08-15 Interactions between semaphorins and plexin–neuropilin receptor complexes in the membranes of live cells Christie, Shaun M. Hao, Jing Tracy, Erin Buck, Matthias Yu, Jennifer S. Smith, Adam W. J Biol Chem Research Article Signaling of semaphorin ligands via their plexin–neuropilin receptors is involved in tissue patterning in the developing embryo. These proteins play roles in cell migration and adhesion but are also important in disease etiology, including in cancer angiogenesis and metastasis. While some structures of the soluble domains of these receptors have been determined, the conformations of the full-length receptor complexes are just beginning to be elucidated, especially within the context of the plasma membrane. Pulsed-interleaved excitation fluorescence cross-correlation spectroscopy allows direct insight into the formation of protein–protein interactions in the membranes of live cells. Here, we investigated the homodimerization of neuropilin-1 (Nrp1), plexin A2, plexin A4, and plexin D1 using pulsed-interleaved excitation fluorescence cross-correlation spectroscopy. Consistent with previous studies, we found that Nrp1, plexin A2, and plexin A4 are present as dimers in the absence of exogenous ligand. Plexin D1, on the other hand, was monomeric under similar conditions, which had not been previously reported. We also found that plexin A2 and A4 assemble into a heteromeric complex. Stimulation with semaphorin 3A or semaphorin 3C neither disrupts nor enhances the dimerization of the receptors when expressed alone, suggesting that activation involves a conformational change rather than a shift in the monomer–dimer equilibrium. However, upon stimulation with semaphorin 3C, plexin D1 and Nrp1 form a heteromeric complex. This analysis of interactions provides a complementary approach to the existing structural and biochemical data that will aid in the development of new therapeutic strategies to target these receptors in cancer. American Society for Biochemistry and Molecular Biology 2021-07-13 /pmc/articles/PMC8350011/ /pubmed/34270956 http://dx.doi.org/10.1016/j.jbc.2021.100965 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Christie, Shaun M.
Hao, Jing
Tracy, Erin
Buck, Matthias
Yu, Jennifer S.
Smith, Adam W.
Interactions between semaphorins and plexin–neuropilin receptor complexes in the membranes of live cells
title Interactions between semaphorins and plexin–neuropilin receptor complexes in the membranes of live cells
title_full Interactions between semaphorins and plexin–neuropilin receptor complexes in the membranes of live cells
title_fullStr Interactions between semaphorins and plexin–neuropilin receptor complexes in the membranes of live cells
title_full_unstemmed Interactions between semaphorins and plexin–neuropilin receptor complexes in the membranes of live cells
title_short Interactions between semaphorins and plexin–neuropilin receptor complexes in the membranes of live cells
title_sort interactions between semaphorins and plexin–neuropilin receptor complexes in the membranes of live cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8350011/
https://www.ncbi.nlm.nih.gov/pubmed/34270956
http://dx.doi.org/10.1016/j.jbc.2021.100965
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