Immunoanalogue of vertebrate beta-adrenergic receptor in the unicellular eukaryote Paramecium

Histochem J. 2002 Jan-Feb;34(1-2):51-6. doi: 10.1023/a:1021399926461.

Abstract

Cell fractionation, SDS-PAGE, quantitative Western blot, confocal immunolocalization and immunogold labelling were performed to find an interpretation of the physiological response of the unicellular eukaryote Paramecium to beta-adrenergic ligands. The 69 kDa polypeptide separated by SDS-PAGE in S2 and P2 Paramecium subcellular fractions cross-reacted with antibody directed against human beta2-adrenergic receptor. This was detected by Western blotting followed by chemiluminescent detection. Quantitative image analysis showed that beta-selective adrenergic agonist (-)-isoproterenol--previously shown to enhance phagocytic activity--evoked redistribution of the adrenergic receptor analogue from membraneous (P2) to cytosolic (S2) fraction. The relative increase in immunoreactive band intensity in S2 reached 80% and was paralleled by a 59% decrease in P2 fraction. Confocal immunofluorescence revealed beta2-adrenergic receptor sites on the cell surface and at the ridge of the cytopharynx--where nascent phagosomes are formed. This localization was confirmed by immunoelectron microscopy. These results indicate that the 69 kDa Paramecium polypeptide immunorelated to vertebrate beta2-adrenergic receptor appeared in this evolutionary ancient cell as a nutrient receptor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Blotting, Western
  • Cell Fractionation
  • Cell Membrane / chemistry
  • Cell Membrane / immunology
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Isoproterenol / pharmacology
  • Microscopy, Confocal
  • Microscopy, Immunoelectron
  • Molecular Weight
  • Paramecium / immunology
  • Paramecium / metabolism*
  • Paramecium / ultrastructure
  • Receptors, Adrenergic, beta-2 / immunology
  • Receptors, Adrenergic, beta-2 / isolation & purification
  • Receptors, Adrenergic, beta-2 / metabolism*
  • Transport Vesicles / chemistry
  • Transport Vesicles / immunology
  • Vertebrates / immunology
  • Vertebrates / metabolism

Substances

  • Receptors, Adrenergic, beta-2
  • Isoproterenol