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Related Experiment Videos

Outer-arm dynein from trout spermatozoa: substructural organization.

S M King1, J L Gatti, A G Moss

  • 1Cell Biology Group, Worcester Foundation for Experimental Biology, Shrewsbury, Massachusetts.

Cell Motility and the Cytoskeleton
|January 1, 1990
PubMed
Summary

Trout sperm outer-arm dynein dissociation revealed distinct subunits, including adenosine triphosphatase (ATPase) complexes and light chains. Heavy chain analysis confirmed molecular masses and evolutionary conservation of intermediate chains.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Outer-arm dynein is a crucial motor protein in sperm flagella, responsible for motility.
  • Understanding dynein's structure and subunit interactions is key to deciphering its function.

Purpose of the Study:

  • To dissociate trout sperm outer-arm dynein and characterize its constituent subunits.
  • To investigate the substructural features and molecular masses of dynein heavy chains.
  • To explore evolutionary conservation of dynein intermediate chains.

Main Methods:

  • Low-ionic-strength dialysis to disrupt dynein.
  • Sucrose density-gradient centrifugation for subunit separation.
  • Vanadate-mediated photolysis to analyze heavy chain fragments.

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  • Immunological analysis using monoclonal antibodies.

    • Main Results:

    • Disruption yielded discrete particles: a 17.5S ATPase, a 9.5S complex, and a 4S light chain (LC 5).
    • Further dissociation produced a 13.1S ATPase containing the beta-heavy chain and intermediate chains (ICs) 3-5.
    • Heavy chain photolysis determined alpha- and beta-heavy chain masses at 430,000 and 415,000 daltons, respectively.
    • Trout IC 2 shares epitopes with Chlamydomonas ICs, indicating conserved sequences.

    Conclusions:

    • The study elucidates the subunit composition and organization of trout sperm outer-arm dynein.
    • The findings provide insights into the intermolecular associations within dynein.
    • Evidence of conserved intermediate chain sequences suggests functional importance and evolutionary stability.