Alterations in the protein pattern of subcellular fractions isolated from Paramecium cells suppressed in phagocytosis

Folia Histochem Cytobiol. 2001;39(4):301-5.

Abstract

SDS-PAGE and quantitative densitometric analysis revealed alterations in the protein pattern of subcellular fractions (100,000 x g) isolated from Paramecium aurelia (299s axenic) cells suppressed in phagocytosis as compared with the control. Two different agents were used to block phagocytosis: the beta-adrenergic antagonist-1-propranolol (200 microM) and inhibitor of calmodulin-dependent processes--trifluoperazine (20 microM). More than 40 polypeptides were identified in the cytosolic (soluble) fractions S1 and S2. A considerable decrease in band intensity was found for three polypeptides: by 60% for 87 kDa band, 52% for 75 kDa and 37% for 42 kDa in comparison to the control, when S2 fractions from propranolol-treated cells of equal load were quantified. TFP treatment evoked only a small decrease in the intensity of the same bands: 9%, 10% and 6%, respectively. The 42 kDa band was identified by Western blot analysis and chemiluminiscent detection to be actin. This result suggests that actin may be a primary target of pharmacological agents used in this study to inhibit Paramecium phagocytic activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / analysis*
  • Adrenergic beta-Antagonists / pharmacology
  • Animals
  • Densitometry
  • Electrophoresis, Polyacrylamide Gel
  • Paramecium / chemistry*
  • Paramecium / drug effects
  • Paramecium / ultrastructure
  • Phagocytosis / drug effects*
  • Propranolol / pharmacology
  • Protozoan Proteins / analysis*
  • Subcellular Fractions
  • Trifluoperazine / pharmacology

Substances

  • Actins
  • Adrenergic beta-Antagonists
  • Protozoan Proteins
  • Trifluoperazine
  • Propranolol