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Abstract:

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.

Registro:

Documento: Artículo
Título:Endocytosis and intracellular dissociation rates of human insulin-insulin receptor complexes by quantum dots in living cells
Autor:Giudice, J.; Jares-Erijman, E.A.; Leskow, F.C.
Filiación:Departamento de Química Biológica, IQUIBICEN-CONICET, Ciudad Universitaria, C1428EGA, Buenos Aires, Argentina
Departamento de Química Orgánica, CIHIDECAR-CONICET, Ciudad Universitaria, C1428EGA, Buenos Aires, Argentina
Departamento de Ciencias Básicas, Universidad Nacional de Luján, Buenos Aires, Argentina
Department of Pathology and Immunology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, United States
Palabras clave: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
Año:2013
Volumen:24
Número:3
Página de inicio:431
Página de fin:442
DOI: http://dx.doi.org/10.1021/bc300526d
Título revista:Bioconjugate Chemistry
Título revista abreviado:Bioconjugate Chem.
ISSN:10431802
CODEN:BCCHE
CAS:streptavidin, 9013-20-1; Antigens, CD; INSR protein, human, 2.7.10.1; Receptor, Insulin, 2.7.10.1
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10431802_v24_n3_p431_Giudice

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Citas:

---------- APA ----------
Giudice, J., Jares-Erijman, E.A. & Leskow, F.C. (2013) . Endocytosis and intracellular dissociation rates of human insulin-insulin receptor complexes by quantum dots in living cells. Bioconjugate Chemistry, 24(3), 431-442.
http://dx.doi.org/10.1021/bc300526d
---------- CHICAGO ----------
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" . Bioconjugate Chemistry 24, no. 3 (2013) : 431-442.
http://dx.doi.org/10.1021/bc300526d
---------- MLA ----------
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" . Bioconjugate Chemistry, vol. 24, no. 3, 2013, pp. 431-442.
http://dx.doi.org/10.1021/bc300526d
---------- VANCOUVER ----------
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. Bioconjugate Chem. 2013;24(3):431-442.
http://dx.doi.org/10.1021/bc300526d