Real-time visual detection of Pi released by flagellar dynein ATPase View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1999-03

AUTHORS

G. G. Vernon, M. Brune, M. R. Webb, D. M. Woolley

ABSTRACT

Using a novel fluorescent probe for Pi, a method for the direct visualization of Pi release from reactivated flagellar dynein ATPase has been developed. The probe undergoes a fluorescence increase when it binds Pi. The technique involves simultaneous imaging of demembranated sperm tails by epi-fluorescence and dark-field microscopy, and the use of the caged ATP technique for axoneme reactivation. To limit diffusion and thus maintain the released Pi within the observed field of view, the assay is carried out within a minute droplet under oil (volume 5-15 pl). The video output of a recursively filtered ICCD camera is used to visualize the fluorescence signal, which is subsequently digitized and automatically analysed on a PC. A major advantage of this technique is that it enables simultaneous analysis of the ATP-utilization rate and the motility of the reactivated axonemes. More... »

PAGES

771-775

References to SciGraph publications

  • 1994-10. The droplet technique: measurement of calcium extrusion from single isolated mammalian cells in PFLÜGERS ARCHIV - EUROPEAN JOURNAL OF PHYSIOLOGY
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/s004240050844

    DOI

    http://dx.doi.org/10.1007/s004240050844

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1046189511

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/10087156


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