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Related Experiment Videos

Preliminary characterization of synaptic vesicle/calmodulin interaction.

N Moskowitz, W Schook, K Beckenstein

    Brain Research
    |March 21, 1983
    PubMed
    Summary

    Calmodulin binds to brain synaptic vesicles at high and low affinities. This binding involves specific proteins, including a Ca2+/calmodulin-dependent protein kinase, influencing key vesicle functions.

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    Area of Science:

    • Neuroscience
    • Molecular Biology
    • Biochemistry

    Background:

    • Calmodulin is a crucial calcium-binding protein involved in cellular signaling.
    • Synaptic vesicles are essential for neurotransmitter release in neurons.

    Purpose of the Study:

    • To investigate the association of calmodulin with brain synaptic vesicle proteins.
    • To identify calmodulin-binding proteins within synaptic vesicles.
    • To determine the functional impact of calmodulin on synaptic vesicle activities.

    Main Methods:

    • Scatchard analysis of radiolabeled calmodulin binding to synaptic vesicles.
    • Affinity chromatography using Sepharose-4B-CNBr-calmodulin to isolate binding proteins.
    • Solubilization of synaptic vesicle proteins with Triton X-100.

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  • Protein identification via molecular weight estimation (MrS) and phosphorylation assays.

    • Main Results:

    • Two binding sites for calmodulin on synaptic vesicles were identified: one high-affinity (Kd = 1.0 nM) and one low-affinity (Kd = 177 nM).
    • Major calmodulin-binding proteins in synaptic vesicles were identified as two doublets (approx. 55 K and 30 K MrS), with minor singlet polypeptides at 62 K and 40 K MrS.
    • Calmodulin was found to stimulate endogenous synaptic vesicle protein kinase, Ca2+, Mg2+-ATPase, and Ca2+ uptake activities.
    • Evidence suggests the synaptic vesicle Ca2+/calmodulin-dependent protein kinase is a 30 K Mr protein.

    Conclusions:

    • Calmodulin interacts with specific proteins in brain synaptic vesicles, modulating their function.
    • The identified calmodulin-binding proteins play roles in regulating synaptic vesicle enzymatic activities and calcium transport.
    • This study elucidates the molecular interactions of calmodulin within the synaptic vesicle.