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Related Experiment Videos

Ligands for insulin receptor isolation.

F M Finn, G Titus, K Hofmann

    Biochemistry
    |June 5, 1984
    PubMed
    Summary
    This summary is machine-generated.

    Researchers developed biotinylated insulins to isolate insulin receptors. They found that the linker length and type, like biotin or dethiobiotin, affect binding and dissociation rates for potential receptor isolation applications.

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    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Endocrinology

    Background:

    • Biotinylated insulins are bivalent molecules with potential for insulin receptor isolation.
    • Simultaneous binding to both insulin receptors and avidin is crucial for effective ligands.
    • Developing biotinylated insulins with simultaneous binding capacity is the focus.

    Purpose of the Study:

    • To create biotinylated and dethiobiotinylated insulins for simultaneous insulin receptor and avidin binding.
    • To investigate the effect of linker length and type on ligand-avidin complex stability.
    • To assess the affinity of these modified insulins for insulin receptors.

    Main Methods:

    • Synthesis of biotinylated and dethiobiotinylated insulins with varying linker lengths (7-20 atoms).

    Related Experiment Videos

  • Complex formation with succinoylavidin and determination of dissociation rates (K-1) using a [14C]biotin exchange assay.
  • Assessment of insulin receptor affinity by measuring 14CO2 formation from [1-14C]glucose in rat adipocytes.

    • Main Results:

    • Biotinylated insulins formed complexes with succinoylavidin, with dissociation rates influenced by linker length and type.
    • Dethiobiotin derivatives showed faster dissociation than biotin derivatives; longer spacer arms decreased dissociation rates.
    • Ligand affinity for insulin receptors correlated with their ability to stimulate glucose metabolism.

    Conclusions:

    • Modified biotinylated insulins can bind simultaneously to insulin receptors and avidin.
    • Linker characteristics significantly impact the stability of biotinylated insulin-avidin complexes.
    • These findings provide a basis for developing novel ligands for insulin receptor isolation and study.