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Decrease by cycloheximide of calcium binding and nonesterified fatty acids in rat-intestinal Golgi-enriched membrane

A Dasmahapatra, J R Walters, M M Weiser

    Biochimica Et Biophysica Acta
    |February 28, 1985
    PubMed
    Summary
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    Vitamin D deficiency impacts calcium (Ca2+) binding in rat intestinal membranes. Protein synthesis inhibition by cycloheximide reduces Ca2+ binding, suggesting a role for newly synthesized proteins in this process.

    Area of Science:

    • Cell Biology
    • Nutritional Biochemistry
    • Gastroenterology

    Background:

    • Intestinal membrane fractions exhibit varying capacities for calcium (Ca2+) binding, with Golgi-enriched fractions showing higher affinity.
    • Vitamin D deficiency is known to impair intestinal Ca2+ absorption.
    • The precise molecular mechanisms underlying Ca2+ binding in intestinal membranes are not fully elucidated.

    Purpose of the Study:

    • To investigate the effect of inhibiting protein synthesis on Ca2+ binding in rat intestinal Golgi-enriched membrane fractions.
    • To explore the role of cycloheximide, a protein synthesis inhibitor, in modulating Ca2+ uptake and transport.
    • To identify potential molecular components, such as fatty acids, involved in Ca2+ binding.

    Main Methods:

    • Preparation of Golgi-enriched, basolateral, and microvillus-enriched membrane fractions from rat intestines.

    Related Experiment Videos

  • Measurement of Ca2+ binding capacity in these fractions.
  • Administration of cycloheximide to rats and assessment of its effects on protein synthesis ([3H]leucine incorporation) and Ca2+ binding.
  • Analysis of nonesterified fatty acids and total lipid phosphate in membrane fractions.

    • Main Results:

    • Golgi-enriched membrane fractions demonstrated higher Ca2+ binding compared to other fractions.
    • Cycloheximide significantly reduced protein synthesis and Ca2+ binding in Golgi membranes.
    • Ca2+ transport across gut sacs was also diminished following cycloheximide treatment.
    • A decrease in nonesterified fatty acids, but not total lipid phosphate, correlated with reduced Ca2+ binding.

    Conclusions:

    • Inhibition of protein synthesis by cycloheximide impairs Ca2+ binding in rat intestinal Golgi membranes.
    • The reduction in Ca2+ binding appears to be mediated by a decrease in nonesterified fatty acids.
    • These findings suggest that newly synthesized proteins are crucial for maintaining the presence of nonesterified fatty acids involved in Ca2+ binding within Golgi membranes.