Histochemistry and Cell Biology

, Volume 105, Issue 4, pp 269–281 | Cite as

Annexins inParamecium cells

Involvement in site-specific positioning of secretory organelles
  • Michael Knochel
  • Roland Kissmehl
  • Johannes-Dieter Wissmann
  • Massoud Momavezi
  • Joachim Hentschel
  • Helmut Plattner
  • Robert D. Burgoyne
Original Paper
Accepted:

Abstract

Annexins were isolated fromParamecium cell homogenates by standard ethylene glycol tetraacetic acid (EGTA) extraction and 100 000-g centrifugation. Two different antibodies (Abs) against synthetic peptides were used, Call-15 and B15, which in mammalian cells recognize a sequence of annexin II or a common sequence occurring in several annexins (except for annexin II), respectively. With anti-Call-15 Abs, western blots from EGTA extracts showed strongly reactive bands of 44.5 and 46 kDa and of higher values. Some of these bands bound to the 100 000-g pellet fraction when Ca2+ was added. Immuno- and affinity labelling revealed selective. Ca2+-dependent labelling of the cell cortex, with enrichent around trichocyst docking sites (facing subplasmalemmal Ca2+ stores). Cortical fluorescence labelling decreased in wild-type (7S) cells when trichocyst ghosts were detached after synchronous exocytosis. Similarly, cortical labelling was reduced when intact trichocysts were detached from the cell surface of non-discharge mutant cells (nd9–28°C, showing identical bands on blots), which then contained numerous heavily labelled phagolysosomes. This strongly suggests annexin downregulation. All together, the dynamic labelling of cortical structures we observed strongly supports involvement of calpactin-like annexins in trichocyst docking. Anti-B15 Abs recognized a band of 51 kDa and some of higher values. These Abs selectively labelled the outlines of the cytoproct, the site of spent phagolysosome exocytosis. In conclusion, our data indicate involvement of specific sets of annexins in site-specific positioning and attachment of widely different secretory organelles at the cell surface inParamecium cells.

Keywords

Synthetic Peptide Fluorescence Labelling Cell Homogenate Docking Site Cell Cortex 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • Michael Knochel
    • 1
  • Roland Kissmehl
    • 1
  • Johannes-Dieter Wissmann
    • 1
  • Massoud Momavezi
    • 1
  • Joachim Hentschel
    • 1
  • Helmut Plattner
    • 1
  • Robert D. Burgoyne
    • 2
  1. 1.Faculty of BiologyUniversity of KonstanzKonstanzGermany
  2. 2.Physiological LaboratoryLiverpoolUK

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