Endocytosis and intracellular dissociation rates of human insulin-insulin receptor complexes by quantum dots in living cells
Insulin signaling is involved in glucose metabolism, cellular growth, and differentiation. Its function is altered in diabetes and many cancer types. Insulin binding to insulin receptor (IR) triggers diverse signaling pathways. However, signal transduction by IR is not mediated exclusively at the ce...
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| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_10431802_v24_n3_p431_Giudice |
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todo:paper_10431802_v24_n3_p431_Giudice2023-10-03T15:58:11Z Endocytosis and intracellular dissociation rates of human insulin-insulin receptor complexes by quantum dots in living cells Giudice, J. Jares-Erijman, E.A. Leskow, F.C. cyan fluorescent protein green fluorescent protein insulin receptor quantum dot recombinant human insulin streptavidin yellow fluorescent protein animal cell article biotinylation cell membrane chemical modification controlled study dissociation drug conjugation endocytosis female human human cell immunofluorescence insulin binding internalization nonhuman phosphorylation plasmid protein folding Western blotting Amino Acid Sequence Animals Antigens, CD Cercopithecus aethiops COS Cells Endocytosis HEK293 Cells HeLa Cells Hep G2 Cells Humans Intracellular Fluid Molecular Sequence Data Quantum Dots Receptor, Insulin Insulin signaling is involved in glucose metabolism, cellular growth, and differentiation. Its function is altered in diabetes and many cancer types. Insulin binding to insulin receptor (IR) triggers diverse signaling pathways. However, signal transduction by IR is not mediated exclusively at the cell surface. Activated ligand-receptor complexes are internalized into endosomes from which the IR recruits adapters acting on substrates that are distinct from those accessible at the membrane. We report the biotinylation of human-recombinant insulin (rhIns) specifically at the position 29 of the B chain. We combined visible fluorescent proteins fused to IR and biotinylated rhIns conjugated with streptavidin-quantum dots to perform extended, quantitative experiments in real time. Modified rhIns bound to the IR and conjugated with the quantum dots was internalized with a rate constant (k) of 0.009 min-1. Dissociation of insulin-IR complex in endocytosed vesicles occurred with k = 0.006 min-1. © 2013 American Chemical Society. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_10431802_v24_n3_p431_Giudice |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
cyan fluorescent protein green fluorescent protein insulin receptor quantum dot recombinant human insulin streptavidin yellow fluorescent protein animal cell article biotinylation cell membrane chemical modification controlled study dissociation drug conjugation endocytosis female human human cell immunofluorescence insulin binding internalization nonhuman phosphorylation plasmid protein folding Western blotting Amino Acid Sequence Animals Antigens, CD Cercopithecus aethiops COS Cells Endocytosis HEK293 Cells HeLa Cells Hep G2 Cells Humans Intracellular Fluid Molecular Sequence Data Quantum Dots Receptor, Insulin |
| spellingShingle |
cyan fluorescent protein green fluorescent protein insulin receptor quantum dot recombinant human insulin streptavidin yellow fluorescent protein animal cell article biotinylation cell membrane chemical modification controlled study dissociation drug conjugation endocytosis female human human cell immunofluorescence insulin binding internalization nonhuman phosphorylation plasmid protein folding Western blotting Amino Acid Sequence Animals Antigens, CD Cercopithecus aethiops COS Cells Endocytosis HEK293 Cells HeLa Cells Hep G2 Cells Humans Intracellular Fluid Molecular Sequence Data Quantum Dots Receptor, Insulin Giudice, J. Jares-Erijman, E.A. Leskow, F.C. Endocytosis and intracellular dissociation rates of human insulin-insulin receptor complexes by quantum dots in living cells |
| topic_facet |
cyan fluorescent protein green fluorescent protein insulin receptor quantum dot recombinant human insulin streptavidin yellow fluorescent protein animal cell article biotinylation cell membrane chemical modification controlled study dissociation drug conjugation endocytosis female human human cell immunofluorescence insulin binding internalization nonhuman phosphorylation plasmid protein folding Western blotting Amino Acid Sequence Animals Antigens, CD Cercopithecus aethiops COS Cells Endocytosis HEK293 Cells HeLa Cells Hep G2 Cells Humans Intracellular Fluid Molecular Sequence Data Quantum Dots Receptor, Insulin |
| description |
Insulin signaling is involved in glucose metabolism, cellular growth, and differentiation. Its function is altered in diabetes and many cancer types. Insulin binding to insulin receptor (IR) triggers diverse signaling pathways. However, signal transduction by IR is not mediated exclusively at the cell surface. Activated ligand-receptor complexes are internalized into endosomes from which the IR recruits adapters acting on substrates that are distinct from those accessible at the membrane. We report the biotinylation of human-recombinant insulin (rhIns) specifically at the position 29 of the B chain. We combined visible fluorescent proteins fused to IR and biotinylated rhIns conjugated with streptavidin-quantum dots to perform extended, quantitative experiments in real time. Modified rhIns bound to the IR and conjugated with the quantum dots was internalized with a rate constant (k) of 0.009 min-1. Dissociation of insulin-IR complex in endocytosed vesicles occurred with k = 0.006 min-1. © 2013 American Chemical Society. |
| format |
JOUR |
| author |
Giudice, J. Jares-Erijman, E.A. Leskow, F.C. |
| author_facet |
Giudice, J. Jares-Erijman, E.A. Leskow, F.C. |
| author_sort |
Giudice, J. |
| title |
Endocytosis and intracellular dissociation rates of human insulin-insulin receptor complexes by quantum dots in living cells |
| title_short |
Endocytosis and intracellular dissociation rates of human insulin-insulin receptor complexes by quantum dots in living cells |
| title_full |
Endocytosis and intracellular dissociation rates of human insulin-insulin receptor complexes by quantum dots in living cells |
| title_fullStr |
Endocytosis and intracellular dissociation rates of human insulin-insulin receptor complexes by quantum dots in living cells |
| title_full_unstemmed |
Endocytosis and intracellular dissociation rates of human insulin-insulin receptor complexes by quantum dots in living cells |
| title_sort |
endocytosis and intracellular dissociation rates of human insulin-insulin receptor complexes by quantum dots in living cells |
| url |
http://hdl.handle.net/20.500.12110/paper_10431802_v24_n3_p431_Giudice |
| work_keys_str_mv |
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| _version_ |
1782025646138982400 |