Characterization of the altered oligosaccharide composition of the insulin receptor on neural-derived cells

Akira Ota; Joshua Shemer; Rebecca M. Pruss; William L. Lowe; Derek LeRoith

doi:10.1016/0006-8993(88)91592-2

Characterization of the altered oligosaccharide composition of the insulin receptor on neural-derived cells

Akira Ota, Joshua Shemer, Rebecca M. Pruss, William L. Lowe, Derek LeRoith^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

12 Scopus citations

Abstract

Typical insulin receptors are present on neuroblastoma cell lines. High affinity binding for insulin was present in membrane preparations from NG108 (a hybrid mouse neuroblastoma-rat glioma) as well as in membranes from SK-N-MC and SK-N-SH, two human neuroblastoma cell lines. Specific [¹²⁵I]insulin binding was 24.4% for NG108, 16.9% for SK-N-MC and 5.2% for SK-N-SH at membrane protein concentrations of 0.4 mg/ml. IC₅₀ for [¹²⁵I]insulin binding was 3.4 nM in NG108 membrane preparations and 0.9 nM for SK-N-SH and 1.8 nM in SK-N-MC membranes. Apparent mol. wt. for the α subunits (identified by specific immunoprecipitation using the anti-insulin receptor antiserum B₁₀) on SDS PAGE was 134 kDa for NG108; 124 kDa for SK-N-MC and 120 kDa for SK-N-SH. Neuraminidase digestion increased the mobility of the α subunit from both NG108 and SK-N-MC receptors to 120 kDa, whereas that from SK-N-SH were unaffected. Endoglycosidase H and endoglycosidase F digestions increased the mobility of the α subunits of all 3 cell lines to varying degrees, suggesting the presence of N-linked glycosy;ation. Insulin induced autophosphorylation of the insulin receptor β subunit in WGA-purified membranes from all 3 cell lines. In addition, phosphorylation of a protein with an apparent mol. wt. 105 kDa was stimulated by insulin in WGA purified membranes from NG108. Tyrosine-specific kinase activity was present in the membranes from each cell line and was stimulated by insulin in a dose-dependent manner from 10^-9 to 10^-6M. Proinsulin was about 100 times less potent in stimulating phosphorylation of the artificial substrate poly (Glu, Tyr)4:1 when compared to insulin in accordance with its lower binding affinity to the insulin receptor. Hexose transport was stimulated by insulin in all 3 cell lines. These results indicate that neuroblastoma cells contain specific insulin receptors and that they may be useful as models for studying the role of insulin in nervous tissue.

Original language	English
Pages (from-to)	1-11
Number of pages	11
Journal	Brain Research
Volume	443
Issue number	1-2
DOIs	https://doi.org/10.1016/0006-8993(88)91592-2
State	Published - 8 Mar 1988
Externally published	Yes

Keywords

Insulin receptor
Neural-derived cell

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/0006-8993(88)91592-2

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@article{e52f77e5236744b594f5f16ccaaaadba,

title = "Characterization of the altered oligosaccharide composition of the insulin receptor on neural-derived cells",

abstract = "Typical insulin receptors are present on neuroblastoma cell lines. High affinity binding for insulin was present in membrane preparations from NG108 (a hybrid mouse neuroblastoma-rat glioma) as well as in membranes from SK-N-MC and SK-N-SH, two human neuroblastoma cell lines. Specific [125I]insulin binding was 24.4% for NG108, 16.9% for SK-N-MC and 5.2% for SK-N-SH at membrane protein concentrations of 0.4 mg/ml. IC50 for [125I]insulin binding was 3.4 nM in NG108 membrane preparations and 0.9 nM for SK-N-SH and 1.8 nM in SK-N-MC membranes. Apparent mol. wt. for the α subunits (identified by specific immunoprecipitation using the anti-insulin receptor antiserum B10) on SDS PAGE was 134 kDa for NG108; 124 kDa for SK-N-MC and 120 kDa for SK-N-SH. Neuraminidase digestion increased the mobility of the α subunit from both NG108 and SK-N-MC receptors to 120 kDa, whereas that from SK-N-SH were unaffected. Endoglycosidase H and endoglycosidase F digestions increased the mobility of the α subunits of all 3 cell lines to varying degrees, suggesting the presence of N-linked glycosy;ation. Insulin induced autophosphorylation of the insulin receptor β subunit in WGA-purified membranes from all 3 cell lines. In addition, phosphorylation of a protein with an apparent mol. wt. 105 kDa was stimulated by insulin in WGA purified membranes from NG108. Tyrosine-specific kinase activity was present in the membranes from each cell line and was stimulated by insulin in a dose-dependent manner from 10-9 to 10-6M. Proinsulin was about 100 times less potent in stimulating phosphorylation of the artificial substrate poly (Glu, Tyr)4:1 when compared to insulin in accordance with its lower binding affinity to the insulin receptor. Hexose transport was stimulated by insulin in all 3 cell lines. These results indicate that neuroblastoma cells contain specific insulin receptors and that they may be useful as models for studying the role of insulin in nervous tissue.",

keywords = "Insulin receptor, Neural-derived cell",

author = "Akira Ota and Joshua Shemer and Pruss, {Rebecca M.} and Lowe, {William L.} and Derek LeRoith",

note = "Funding Information: We wish to thank Drs. Simeon Taylor and Jesse Roth for their advice regarding these experiments and Mrs. Violet Katz for secretarial assistance. This work was supported in part by a grant from the Washington D.C. Affiliation of the American Diabetes Association.",

year = "1988",

month = mar,

day = "8",

doi = "10.1016/0006-8993(88)91592-2",

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volume = "443",

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journal = "Brain Research",

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T1 - Characterization of the altered oligosaccharide composition of the insulin receptor on neural-derived cells

AU - Ota, Akira

AU - Shemer, Joshua

AU - Pruss, Rebecca M.

AU - Lowe, William L.

AU - LeRoith, Derek

N1 - Funding Information: We wish to thank Drs. Simeon Taylor and Jesse Roth for their advice regarding these experiments and Mrs. Violet Katz for secretarial assistance. This work was supported in part by a grant from the Washington D.C. Affiliation of the American Diabetes Association.

PY - 1988/3/8

Y1 - 1988/3/8

N2 - Typical insulin receptors are present on neuroblastoma cell lines. High affinity binding for insulin was present in membrane preparations from NG108 (a hybrid mouse neuroblastoma-rat glioma) as well as in membranes from SK-N-MC and SK-N-SH, two human neuroblastoma cell lines. Specific [125I]insulin binding was 24.4% for NG108, 16.9% for SK-N-MC and 5.2% for SK-N-SH at membrane protein concentrations of 0.4 mg/ml. IC50 for [125I]insulin binding was 3.4 nM in NG108 membrane preparations and 0.9 nM for SK-N-SH and 1.8 nM in SK-N-MC membranes. Apparent mol. wt. for the α subunits (identified by specific immunoprecipitation using the anti-insulin receptor antiserum B10) on SDS PAGE was 134 kDa for NG108; 124 kDa for SK-N-MC and 120 kDa for SK-N-SH. Neuraminidase digestion increased the mobility of the α subunit from both NG108 and SK-N-MC receptors to 120 kDa, whereas that from SK-N-SH were unaffected. Endoglycosidase H and endoglycosidase F digestions increased the mobility of the α subunits of all 3 cell lines to varying degrees, suggesting the presence of N-linked glycosy;ation. Insulin induced autophosphorylation of the insulin receptor β subunit in WGA-purified membranes from all 3 cell lines. In addition, phosphorylation of a protein with an apparent mol. wt. 105 kDa was stimulated by insulin in WGA purified membranes from NG108. Tyrosine-specific kinase activity was present in the membranes from each cell line and was stimulated by insulin in a dose-dependent manner from 10-9 to 10-6M. Proinsulin was about 100 times less potent in stimulating phosphorylation of the artificial substrate poly (Glu, Tyr)4:1 when compared to insulin in accordance with its lower binding affinity to the insulin receptor. Hexose transport was stimulated by insulin in all 3 cell lines. These results indicate that neuroblastoma cells contain specific insulin receptors and that they may be useful as models for studying the role of insulin in nervous tissue.

AB - Typical insulin receptors are present on neuroblastoma cell lines. High affinity binding for insulin was present in membrane preparations from NG108 (a hybrid mouse neuroblastoma-rat glioma) as well as in membranes from SK-N-MC and SK-N-SH, two human neuroblastoma cell lines. Specific [125I]insulin binding was 24.4% for NG108, 16.9% for SK-N-MC and 5.2% for SK-N-SH at membrane protein concentrations of 0.4 mg/ml. IC50 for [125I]insulin binding was 3.4 nM in NG108 membrane preparations and 0.9 nM for SK-N-SH and 1.8 nM in SK-N-MC membranes. Apparent mol. wt. for the α subunits (identified by specific immunoprecipitation using the anti-insulin receptor antiserum B10) on SDS PAGE was 134 kDa for NG108; 124 kDa for SK-N-MC and 120 kDa for SK-N-SH. Neuraminidase digestion increased the mobility of the α subunit from both NG108 and SK-N-MC receptors to 120 kDa, whereas that from SK-N-SH were unaffected. Endoglycosidase H and endoglycosidase F digestions increased the mobility of the α subunits of all 3 cell lines to varying degrees, suggesting the presence of N-linked glycosy;ation. Insulin induced autophosphorylation of the insulin receptor β subunit in WGA-purified membranes from all 3 cell lines. In addition, phosphorylation of a protein with an apparent mol. wt. 105 kDa was stimulated by insulin in WGA purified membranes from NG108. Tyrosine-specific kinase activity was present in the membranes from each cell line and was stimulated by insulin in a dose-dependent manner from 10-9 to 10-6M. Proinsulin was about 100 times less potent in stimulating phosphorylation of the artificial substrate poly (Glu, Tyr)4:1 when compared to insulin in accordance with its lower binding affinity to the insulin receptor. Hexose transport was stimulated by insulin in all 3 cell lines. These results indicate that neuroblastoma cells contain specific insulin receptors and that they may be useful as models for studying the role of insulin in nervous tissue.

KW - Insulin receptor

KW - Neural-derived cell

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Characterization of the altered oligosaccharide composition of the insulin receptor on neural-derived cells

Abstract

Keywords

UN SDGs

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