Protoplasma

, Volume 168, Issue 3, pp 113–124

Biochemical and immunological characterization of intermicrotubular cement in the feeding apparatus of phagotrophic eugienoids:Entosiphon, Peranema, andPloeotia

  • A. Belhadri
  • Danielle Bayle
  • G. Brugerolle
Article

DOI: 10.1007/BF01666257

Cite this article as:
Belhadri, A., Bayle, D. & Brugerolle, G. Protoplasma (1992) 168: 113. doi:10.1007/BF01666257

Summary

A monoclonal antibody (IIID12) obtained from mice immunized against the entireEntosiphon cytoskeleton highlights the feeding apparatus ofEntosiphon, Peranema, andPloeotia by IF. IGS at the ultrastructural level shows that it labels the cementing material surrounding the microtubular bundles in the three species studied. InEntosiphon additional structures, such as the supplementary plaque, the scaffold structure and the lenticular structure or canal thickening, are also detected by the antibody. Immunoblotting analysis after SDS-PAGE reveals a positive reaction with this antibody to the 58 and 66kDa protein bands inEntosiphon, 82 and 84kDa inPeranema, and 56 and 60kDa inPloeotia. These results demonstrate biochemical homologies in the proteins of the cement material in the three heteronematal eugienoids studied. The possible role of these proteins in microtubule assembly and stabilization is discussed, as well as the role of the cementing material in the mode of the feeding apparatus motion during the ingestion of food.

Keywords

CytoskeletonPhagotrophicEugienoidsImmunochemistryUltrastructure

Abbreviations

BSA

bovine serum albumin

DMSO

dimethyl sulfoxide

EDTA

ethylenediaminetetraacetic acid

IF

immunofluorescence

IGS

immunogold staining

HAT

hypoxantin-aminopterinthymidine

MAb

monoclonal antibody

MEM

minimum essential medium

PBS

phosphate buffered saline

PMSF

phenylmethanesulfonyl fluoride

TAME

Nα-p-tosyl-L-arginine methyl-ester

TLCK

Nα-p-tosyl-L-lysine chloromethyl ketone

SDS-PAGE

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • A. Belhadri
    • 1
  • Danielle Bayle
    • 1
  • G. Brugerolle
    • 1
  1. 1.Laboratoire de Zoologie-ProtistologieUniversité Blaise PascalAubièreFrance