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Axokinin phosphorylation by cAMP-dependent protein kinase is sufficient for activation of sperm flagellar motility.

Publication ,  Journal Article
Tash, JS; Hidaka, H; Means, AR
Published in: J Cell Biol
August 1986

Using a selective inhibitor of cAMP-dependent protein kinase, N-[2(methylamino)ethyl]-5-isoquinolinesulfonamide (H-8), the requirement for cAMP-dependent phosphoproteins in the initiation of dog sperm flagellar motility was examined. H-8 inhibited motility of live as well as reactivated sperm in a dose-dependent manner. The half-maximal inhibition of reactivated motility (32 microM) paralleled the inhibition of pure catalytic subunit of cAMP-dependent protein kinase (50 microM) measured under the same conditions. H-8 inhibited protein phosphorylation both in whole models and in isolated Nonidet P-40 (NP-40) extracts of sperm. Axokinin, the heat-stable NP-40-soluble protein whose phosphorylation is required for flagellar reactivation, represented 97% of the de novo phosphate incorporation in the NP-40 extract after stimulation by cAMP. 500 microM H-8 inhibited axokinin phosphorylation by 87%. When sperm were reactivated in the presence of up to 5 mM H-8 with NP-40 extract that had been prephosphorylated with cAMP-dependent protein kinase, then neither cAMP nor cAMP-dependent protein kinase activity was required for full flagellar reactivation. If sperm were rendered completely immotile by pretreatment with H-8, then the resulting model remained immotile in the continued presence of H-8 unless prephosphorylated axokinin was added. These results suggest that phosphorylated axokinin is not only required for flagellar reactivation but is sufficient as well.

Duke Scholars

Published In

J Cell Biol

DOI

ISSN

0021-9525

Publication Date

August 1986

Volume

103

Issue

2

Start / End Page

649 / 655

Location

United States

Related Subject Headings

  • Thiazolidinediones
  • Spermatozoa
  • Sperm Motility
  • Protein Kinases
  • Protein Kinase Inhibitors
  • Phosphorylation
  • Phosphoproteins
  • Male
  • Dogs
  • Developmental Biology
 

Citation

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Tash, J. S., Hidaka, H., & Means, A. R. (1986). Axokinin phosphorylation by cAMP-dependent protein kinase is sufficient for activation of sperm flagellar motility. J Cell Biol, 103(2), 649–655. https://doi.org/10.1083/jcb.103.2.649
Tash, J. S., H. Hidaka, and A. R. Means. “Axokinin phosphorylation by cAMP-dependent protein kinase is sufficient for activation of sperm flagellar motility.J Cell Biol 103, no. 2 (August 1986): 649–55. https://doi.org/10.1083/jcb.103.2.649.
Tash, J. S., et al. “Axokinin phosphorylation by cAMP-dependent protein kinase is sufficient for activation of sperm flagellar motility.J Cell Biol, vol. 103, no. 2, Aug. 1986, pp. 649–55. Pubmed, doi:10.1083/jcb.103.2.649.

Published In

J Cell Biol

DOI

ISSN

0021-9525

Publication Date

August 1986

Volume

103

Issue

2

Start / End Page

649 / 655

Location

United States

Related Subject Headings

  • Thiazolidinediones
  • Spermatozoa
  • Sperm Motility
  • Protein Kinases
  • Protein Kinase Inhibitors
  • Phosphorylation
  • Phosphoproteins
  • Male
  • Dogs
  • Developmental Biology