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Identification of function-regulating antibodies targeting the receptor protein tyrosine phosphatase sigma ectodomain.

ABSTRACT: Receptor tyrosine phosphatase sigma (RPTP?) plays an important role in the regulation of axonal outgrowth and neural regeneration. Recent studies have identified two RPTP? ligands, chondroitin sulfate proteoglycans (CSPGs) and heparan sulfate proteoglycans (HSPG), which can modulate RPTP? activity by affecting its dimerization status. Here, we developed a split luciferase assay to monitor RPTP? dimerization in living cells. Using this system, we demonstrate that heparin, an analog of heparan sulfate, induced the dimerization of RPTP?, whereas chondroitin sulfate increased RPTP? activity by inhibiting RPTP? dimerization. Also, we generated several novel RPTP? IgG monoclonal antibodies, to identify one that modulates its activity by inducing/stabilizing dimerization in living cells. Lastly, we demonstrate that this antibody promotes neurite outgrowth in SH-SY5Y cells. In summary, we demonstrated that the split luciferase RPTP? activity assay is a novel high-throughput approach for discovering novel RPTP? modulators that can promote axonal outgrowth and neural regeneration.

SUBMITTER: Wu CL

PROVIDER: S-EPMC5449173 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Identification of function-regulating antibodies targeting the receptor protein tyrosine phosphatase sigma ectodomain.

Wu Chia-Lun CL Hardy Serge S Aubry Isabelle I Landry Melissa M Haggarty Allison A Saragovi Horacio Uri HU Tremblay Michel L ML

PloS one 20170530 5

Receptor tyrosine phosphatase sigma (RPTPσ) plays an important role in the regulation of axonal outgrowth and neural regeneration. Recent studies have identified two RPTPσ ligands, chondroitin sulfate proteoglycans (CSPGs) and heparan sulfate proteoglycans (HSPG), which can modulate RPTPσ activity by affecting its dimerization status. Here, we developed a split luciferase assay to monitor RPTPσ dimerization in living cells. Using this system, we demonstrate that heparin, an analog of heparan sul ...[more]

PMID: 28558026

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Project description:Reversible tyrosine phosphorylation, catalyzed by receptor tyrosine kinases and receptor tyrosine phosphatases, plays an essential part in cell signaling during axonal development. Receptor protein tyrosine phosphatase-sigma has been implicated in the growth, guidance and repair of retinal axons. This phosphatase has also been implicated in motor axon growth and innervation. Insect orthologs of receptor protein tyrosine phosphatase-sigma are also implicated in the recognition of muscle target cells. A potential extracellular ligand for vertebrate receptor protein tyrosine phosphatase-sigma has been previously localized in developing skeletal muscle. The identity of this muscle ligand is currently unknown, but it appears to be unrelated to the heparan sulfate ligands of receptor protein tyrosine phosphatase-sigma. In this study, we have used affinity chromatography and tandem MS to identify nucleolin as a binding partner for receptor protein tyrosine phosphatase-sigma in skeletal muscle tissue. Nucleolin, both from tissue lysates and in purified form, binds to receptor protein tyrosine phosphatase-sigma ectodomains. Its expression pattern also overlaps with that of the receptor protein tyrosine phosphatase-sigma-binding partner previously localized in muscle, and nucleolin can also be found in retinal basement membranes. We demonstrate that a significant amount of muscle-associated nucleolin is present on the cell surface of developing myotubes, and that two nucleolin-binding components, lactoferrin and the HB-19 peptide, can block the interaction of receptor protein tyrosine phosphatase-sigma ectodomains with muscle and retinal basement membranes in tissue sections. These data suggest that muscle cell surface-associated nucleolin represents at least part of the muscle binding site for axonal receptor protein tyrosine phosphatase-sigma and that nucleolin may also be a necessary component of basement membrane binding sites of receptor protein tyrosine phosphatase-sigma.

Dataset Information

Identification of function-regulating antibodies targeting the receptor protein tyrosine phosphatase sigma ectodomain.

Publications

Identification of function-regulating antibodies targeting the receptor protein tyrosine phosphatase sigma ectodomain.

Similar Datasets

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