Cryptdin 3 forms anion selective channels in cytoplasmic membranes of human embryonic kidney cells.

Cryptdins are antimicrobial peptides secreted by Paneth cells located at the base of intestinal crypts. In addition to their antimicrobial function, cryptdins may also regulate salt and water secretion by intestinal epithelial cells. Recent work with short-circuit current measurements indicated that at least one cryptdin peptide, cryptdin 3, induces apical conductance(s) in Cl(-) secretory, including cystic fibrosis, epithelia. In the present study, we characterized the cryptdin 3-induced anion channel activity in human embryonic kidney (HEK) cells with single-channel patch-clamp techniques. The patch pipette was filled with solution containing different concentrations of cryptdin 3, and, after gigaseal formation, the channel activity was recorded with either cell-attached or inside-out patch modes. We found an anion selective channel with a conductance of 15 pS and open probability of 0.19, regardless of cryptdin 3 concentration. The mean open and closed times varied with the cryptdin 3 concentration. For cryptdin 3 concentrations of 10, 4, 1, and 0.5 microg/ml in the pipette, the corresponding mean open times were 1.2, 7.0, 9.0, and 17.4 ms and the corresponding mean closed times were 1.1, 1.6, 4.2, and 12.5 ms. These results suggest that cryptdin 3 forms anion-selective channels on the cytoplasmic membrane of HEK cells and that the kinetics of one such channel are affected by its interaction with other such channels.
AuthorsGang Yue, Didier Merlin, Michael E Selsted, Wayne I Lencer, James L Madara, Douglas C Eaton
JournalAmerican journal of physiology. Gastrointestinal and liver physiology (Am J Physiol Gastrointest Liver Physiol) Vol. 282 Issue 5 Pg. G757-65 (May 2002) ISSN: 0193-1857 [Print] United States
PMID11960772 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)
Chemical References
  • Anions
  • Anti-Infective Agents
  • Chloride Channels
  • Defensins
  • Proteins
  • cryptdin 3
  • Amino Acid Sequence
  • Animals
  • Anions (metabolism)
  • Anti-Infective Agents (chemistry, pharmacology)
  • Cell Line
  • Cell Membrane (physiology)
  • Chloride Channels (physiology)
  • Cytoplasm (physiology)
  • Defensins (physiology)
  • Dose-Response Relationship, Drug
  • Humans
  • Ion Channel Gating (drug effects, physiology)
  • Kidney (cytology)
  • Mice
  • Molecular Sequence Data
  • Patch-Clamp Techniques
  • Proteins (chemistry, pharmacology)

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