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Human brain tubulin purification: decrease in soluble tubulin with age.

S C Yan, S Hwang, T D Rustan

    Neurochemical Research
    |January 1, 1985
    PubMed
    Summary
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    Human brain tubulin significantly decreases with age. A novel purification method yields polymerization-competent human tubulin, similar to mammalian tubulin, enabling further research into brain function.

    Area of Science:

    • Neuroscience
    • Biochemistry
    • Cell Biology

    Background:

    • Soluble tubulin concentration in the human cerebral cortex declines significantly with age (75% from birth to 90 years).
    • Postmortem human brain soluble fractions exhibit lower high molecular weight proteins compared to nonhuman species.

    Purpose of the Study:

    • To develop a novel method for purifying polymerization-competent tubulin from human cerebral cortex.
    • To characterize the properties of purified human brain tubulin.

    Main Methods:

    • Utilized [3H]colchicine binding to assess soluble tubulin in the 100,000 g supernatant of human cerebral cortex.
    • Polymerized human brain tubulin into microtubules using DEAE-dextran.
    • Investigated microtubule stability under varying conditions (cold, calcium, glutamate).

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  • Developed and applied a novel purification technique for human brain tubulin.

    • Main Results:

    • Demonstrated a drastic age-related decrease in soluble tubulin in the human cerebral cortex.
    • Showcased the ability to polymerize human brain tubulin into stable microtubules.
    • Identified conditions (1 M glutamate) that induce cold lability and depolymerization.
    • Purified human brain tubulin exhibits properties comparable to mammalian tubulin.

    Conclusions:

    • A novel method effectively purifies polymerization-competent tubulin from human cerebral cortex.
    • Purified human brain tubulin shares biochemical and structural similarities with tubulin from other mammals.
    • This purified tubulin is suitable for studying microtubule dynamics and brain aging.