Disruption of guanylyl cyclase signalling pathway mediating intestinal secretion

S. J. Parkinson, A. E. Alekseev, L. A. Gomez, F. Wagner, S. Crupper, D. C. Robertson, Andre Terzic, S. A. Waldman

Research output: Contribution to journalArticle

Abstract

The present studies establish that an intracellular nucleotide dependent pathway disrupts cyclic GMP (cGMP) production by guanylyl cyclase C (GCC) and guanylyl cyclase A (GCA). Inhibition requires the kinase-homology domain since deletion of this region yields a guanylyl cyclase insensitive to the nucleotide inhibitor, 2-chloro-ATP (2ClATP). Immunopurificd GCA is insensitive to 2ClATP suggesting additional factors are required for inhibition. Diarrhea induced by E. coli heat-stable enterotoxin (STa) is mediated by guanylyl cyclase C (GCC). Incubation of Caco 2 human intestinal epithelial cells with 2-chloroadenosine (2ClAdo) prevented STa dependent cGMP production and associated chloride (Cl-) flux which underlies intestinal secretion. Inhibition was concentration and time dependent correlating with the metabolic conversion of 2ClAdo to 2ClATP. Guanylyl cyclase activity in membranes prepared from 2ClAdo treated cells was inhibited, compared to membranes from control cells, demonstrating a noncompetitive mechanism. Treatment of Caco 2 cells with 2ClAdo also prevented STa-induced Cl- flux. Incubation of 2ClAdo treated cells with the cell permeant analog 8Br-cGMP reconstituted the Cl- current, demonstrating that inhibition of Cl- flux reflected selective disruption of ligand stim-ulation of GCC rather than the chloride channel. Thus, the components required for adenine nucleotide inhibition of receptor guanylyl cyclase signaling are present in intact mammalian cells, establishing the utility of this pathway for elucidating the mechanisms regulating GCC and other receptor guanylyl cyclases.

Original languageEnglish (US)
Pages (from-to)235
Number of pages1
JournalClinical Pharmacology and Therapeutics
Volume61
Issue number2
StatePublished - 1997

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2-Chloroadenosine
Intestinal Secretions
Guanylate Cyclase
Chlorides
Cyclic GMP
Enterotoxins
Guanylate Cyclase-Coupled Receptors
Nucleotides
Chloride Channels
Caco-2 Cells
Adenine Nucleotides
Diarrhea
Phosphotransferases
Epithelial Cells
Cell Membrane
Ligands
Membranes
enterotoxin receptor
2-chloro-ATP

ASJC Scopus subject areas

  • Pharmacology

Cite this

Parkinson, S. J., Alekseev, A. E., Gomez, L. A., Wagner, F., Crupper, S., Robertson, D. C., ... Waldman, S. A. (1997). Disruption of guanylyl cyclase signalling pathway mediating intestinal secretion. Clinical Pharmacology and Therapeutics, 61(2), 235.

Disruption of guanylyl cyclase signalling pathway mediating intestinal secretion. / Parkinson, S. J.; Alekseev, A. E.; Gomez, L. A.; Wagner, F.; Crupper, S.; Robertson, D. C.; Terzic, Andre; Waldman, S. A.

In: Clinical Pharmacology and Therapeutics, Vol. 61, No. 2, 1997, p. 235.

Research output: Contribution to journalArticle

Parkinson, SJ, Alekseev, AE, Gomez, LA, Wagner, F, Crupper, S, Robertson, DC, Terzic, A & Waldman, SA 1997, 'Disruption of guanylyl cyclase signalling pathway mediating intestinal secretion', Clinical Pharmacology and Therapeutics, vol. 61, no. 2, pp. 235.
Parkinson SJ, Alekseev AE, Gomez LA, Wagner F, Crupper S, Robertson DC et al. Disruption of guanylyl cyclase signalling pathway mediating intestinal secretion. Clinical Pharmacology and Therapeutics. 1997;61(2):235.
Parkinson, S. J. ; Alekseev, A. E. ; Gomez, L. A. ; Wagner, F. ; Crupper, S. ; Robertson, D. C. ; Terzic, Andre ; Waldman, S. A. / Disruption of guanylyl cyclase signalling pathway mediating intestinal secretion. In: Clinical Pharmacology and Therapeutics. 1997 ; Vol. 61, No. 2. pp. 235.
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