Cellular uptake and in situ binding of a peptide agonist for calmodulin

Biochem Biophys Res Commun. 2000 Oct 22;277(2):462-9. doi: 10.1006/bbrc.2000.3691.

Abstract

We have used the method of inverted hydropathy to develop peptides that interact with EF hands of calmodulin (CaM). Previously we have shown these peptides specifically interact with their desired target in a productive manner, in that they activated CaM in the absence of Ca(2+). Therefore, we sought to determine whether these peptides would enter cells, remain intact, and interact with CaM in the interior of the cell. Using several techniques we have demonstrated cellular uptake, stability, and an intracellular interaction with CaM with fluorescein-labeled and radiolabeled peptides in Jurkat T cells. The results suggest that these peptides may be useful in the study and the manipulation of Ca(2+)-mediated pathways in cells.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Blotting, Western
  • Calcium / metabolism
  • Calmodulin / agonists*
  • Calmodulin / chemistry*
  • Calmodulin / metabolism
  • Cattle
  • Cells, Cultured
  • Chromatography, High Pressure Liquid
  • Cross-Linking Reagents / pharmacology
  • Dose-Response Relationship, Drug
  • Fluorescein / pharmacology
  • Humans
  • Jurkat Cells
  • Kinetics
  • Microscopy, Fluorescence
  • Peptides / chemistry*
  • Peptides / metabolism
  • Peptides / pharmacokinetics
  • Phosphoric Diester Hydrolases / metabolism
  • Precipitin Tests
  • Protein Binding
  • Signal Transduction
  • Subcellular Fractions
  • Temperature
  • Time Factors

Substances

  • Calmodulin
  • Cross-Linking Reagents
  • Peptides
  • Phosphoric Diester Hydrolases
  • Calcium
  • Fluorescein