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

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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.

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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

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Authors and Affiliations

  1. Laboratoire de Zoologie-Protistologie, Université Blaise Pascal, UA CNRS 138, F-63177, Aubière, France

    A. Belhadri, Danielle Bayle & G. Brugerolle

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  1. A. Belhadri
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  2. Danielle Bayle
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  3. G. Brugerolle
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Belhadri, A., Bayle, D. & Brugerolle, G. Biochemical and immunological characterization of intermicrotubular cement in the feeding apparatus of phagotrophic eugienoids:Entosiphon, Peranema, andPloeotia . Protoplasma 168, 113–124 (1992). https://doi.org/10.1007/BF01666257

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