A 43 kDa protein inserted at the plasma membrane-cytoskeleton interface in rumen entodiniomorphid ciliates |
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Authors: | A Sghir D David M Ferrier |
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Institution: | (1) Biologie Comparée des Protistes, Université de Clermont-Ferrand II, Complexe Scientifique des Cézeaux, F-63177 Aubière Cedex, France |
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Abstract: | Summary The P-43 ofEudiplodinium and homologous proteins in three other entodiniomorphid species, free-living ciliates, flagellates, and HeLa cells, were identified at the plasma membrane-cytoskeleton interface. Proteins cross-reacting with MAb B6 were also located at the ciliary inner surface of the plasma membrane. Due to the strong adhesion of the plasma membrane to the underlying cytoskeleton, classical extraction with detergents, urea, NaOH, and PTA, failed to separate the two components completely. However, the extraction properties of P-43, associated with its membrane-cytoskeleton interactive functions, suggest that this unglycosylated protein may present some analogies with proteins of the intermediate filaments. Their ubiquity and localization suggest that P-43 and MAb B6 crossreacting proteins may not be strictly epiplasmic but could be amphitropic proteins, strongly anchored to both the plasma membrane and the underlying microfilament framework, via protein-protein binding or by direct insertion in the lipid bilayer.Abbreviations BSA
bovine serum albumin
- Con A
concanavalin A
- EDTA
ethylene diamine tetraacetic acid
- EM
electron microscopy
- IF
intermediate filaments
- MAb
monoclonal antibody
- MET
2-mercaptoethanol
- MW
molecular weight
- PAb
polyclonal antibody
- PBS
phosphate buffered saline
- PMSF
phenylmethylsulfonyl fluoride
- PTA
phosphotungstic acid
- SDS
sodiumdodecyl sulfate
- TAME
Na-p-tosyl-arginine methyl ester
- TLCK
Na-p-tosyl-lysine chloromethyl ketone |
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Keywords: | Biochemical properties Cytoskeleton-plasma membrane interface Immunological properties 43 kDa Protein Entodiniomorphid ciliates |
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