Novel tool for the study of cholecystokinin-stimulated pancreatic enzyme secretion

H. Y. Gaisano, U. G. Klueppelberg, D. I. Pinon, M. A. Pfenning, S. P. Powers, Laurence J Miller

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

The molecular events that mediate cholecystokinin (CCK)-stimulated pancreatic secretion are not well defined because of the complex receptor-binding and concentration-response characteristics of this hormone. Functional models of receptor occupancy initiating the cascade leading to secretion have been complicated by the inhibition of secretion effected by supra-maximal concentrations of CCK. Recent report of a CCK analogue that does not exhibit supramaximal inhibition led us to synthesize a similar analogue that could also be radiolabeled for studies of receptor binding and affinity labeling, and for studies of second messenger activity. This probe, D-Tyr-Gly-[(Nle28,31)CCK-26-32]-phenethyl ester, was a fully efficacious secretagogue with no supramaximal inhibition, and, unlike native hormone, bound to a single class of sites present on both acini and membranes. Occupation of this site correlated well with stimulation of secretion. Evidence that this was indeed a CCK-binding site were the abilities of CCK and antagonist L-364,718 to inhibit binding of this analogue. Affinity labeling confirmed the identity of the site mediating secretory stimulation as a M(r) = 85,000-95,000 protein. Whereas the nonhydrolyzable guanosine triphosphate analogue, 5'-guanylyl-imidodiphosphate, was a potent inhibitor of CCK binding, it had no effect on binding of this secretagogue, suggesting that a novel cascade not involving a guanine nucleotide-binding protein mediates CCK stimulation of pancreatic secretion.

Original languageEnglish (US)
Pages (from-to)321-325
Number of pages5
JournalJournal of Clinical Investigation
Volume83
Issue number1
StatePublished - 1989

Fingerprint

Cholecystokinin
Enzymes
Devazepide
Hormones
Guanylyl Imidodiphosphate
Guanine Nucleotides
Second Messenger Systems
Guanosine Triphosphate
Occupations
Carrier Proteins
Esters
Binding Sites
Membranes
Proteins

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Gaisano, H. Y., Klueppelberg, U. G., Pinon, D. I., Pfenning, M. A., Powers, S. P., & Miller, L. J. (1989). Novel tool for the study of cholecystokinin-stimulated pancreatic enzyme secretion. Journal of Clinical Investigation, 83(1), 321-325.

Novel tool for the study of cholecystokinin-stimulated pancreatic enzyme secretion. / Gaisano, H. Y.; Klueppelberg, U. G.; Pinon, D. I.; Pfenning, M. A.; Powers, S. P.; Miller, Laurence J.

In: Journal of Clinical Investigation, Vol. 83, No. 1, 1989, p. 321-325.

Research output: Contribution to journalArticle

Gaisano, HY, Klueppelberg, UG, Pinon, DI, Pfenning, MA, Powers, SP & Miller, LJ 1989, 'Novel tool for the study of cholecystokinin-stimulated pancreatic enzyme secretion', Journal of Clinical Investigation, vol. 83, no. 1, pp. 321-325.
Gaisano HY, Klueppelberg UG, Pinon DI, Pfenning MA, Powers SP, Miller LJ. Novel tool for the study of cholecystokinin-stimulated pancreatic enzyme secretion. Journal of Clinical Investigation. 1989;83(1):321-325.
Gaisano, H. Y. ; Klueppelberg, U. G. ; Pinon, D. I. ; Pfenning, M. A. ; Powers, S. P. ; Miller, Laurence J. / Novel tool for the study of cholecystokinin-stimulated pancreatic enzyme secretion. In: Journal of Clinical Investigation. 1989 ; Vol. 83, No. 1. pp. 321-325.
@article{1a94166b080f410787b26e44b094dc6b,
title = "Novel tool for the study of cholecystokinin-stimulated pancreatic enzyme secretion",
abstract = "The molecular events that mediate cholecystokinin (CCK)-stimulated pancreatic secretion are not well defined because of the complex receptor-binding and concentration-response characteristics of this hormone. Functional models of receptor occupancy initiating the cascade leading to secretion have been complicated by the inhibition of secretion effected by supra-maximal concentrations of CCK. Recent report of a CCK analogue that does not exhibit supramaximal inhibition led us to synthesize a similar analogue that could also be radiolabeled for studies of receptor binding and affinity labeling, and for studies of second messenger activity. This probe, D-Tyr-Gly-[(Nle28,31)CCK-26-32]-phenethyl ester, was a fully efficacious secretagogue with no supramaximal inhibition, and, unlike native hormone, bound to a single class of sites present on both acini and membranes. Occupation of this site correlated well with stimulation of secretion. Evidence that this was indeed a CCK-binding site were the abilities of CCK and antagonist L-364,718 to inhibit binding of this analogue. Affinity labeling confirmed the identity of the site mediating secretory stimulation as a M(r) = 85,000-95,000 protein. Whereas the nonhydrolyzable guanosine triphosphate analogue, 5'-guanylyl-imidodiphosphate, was a potent inhibitor of CCK binding, it had no effect on binding of this secretagogue, suggesting that a novel cascade not involving a guanine nucleotide-binding protein mediates CCK stimulation of pancreatic secretion.",
author = "Gaisano, {H. Y.} and Klueppelberg, {U. G.} and Pinon, {D. I.} and Pfenning, {M. A.} and Powers, {S. P.} and Miller, {Laurence J}",
year = "1989",
language = "English (US)",
volume = "83",
pages = "321--325",
journal = "Journal of Clinical Investigation",
issn = "0021-9738",
publisher = "The American Society for Clinical Investigation",
number = "1",

}

TY - JOUR

T1 - Novel tool for the study of cholecystokinin-stimulated pancreatic enzyme secretion

AU - Gaisano, H. Y.

AU - Klueppelberg, U. G.

AU - Pinon, D. I.

AU - Pfenning, M. A.

AU - Powers, S. P.

AU - Miller, Laurence J

PY - 1989

Y1 - 1989

N2 - The molecular events that mediate cholecystokinin (CCK)-stimulated pancreatic secretion are not well defined because of the complex receptor-binding and concentration-response characteristics of this hormone. Functional models of receptor occupancy initiating the cascade leading to secretion have been complicated by the inhibition of secretion effected by supra-maximal concentrations of CCK. Recent report of a CCK analogue that does not exhibit supramaximal inhibition led us to synthesize a similar analogue that could also be radiolabeled for studies of receptor binding and affinity labeling, and for studies of second messenger activity. This probe, D-Tyr-Gly-[(Nle28,31)CCK-26-32]-phenethyl ester, was a fully efficacious secretagogue with no supramaximal inhibition, and, unlike native hormone, bound to a single class of sites present on both acini and membranes. Occupation of this site correlated well with stimulation of secretion. Evidence that this was indeed a CCK-binding site were the abilities of CCK and antagonist L-364,718 to inhibit binding of this analogue. Affinity labeling confirmed the identity of the site mediating secretory stimulation as a M(r) = 85,000-95,000 protein. Whereas the nonhydrolyzable guanosine triphosphate analogue, 5'-guanylyl-imidodiphosphate, was a potent inhibitor of CCK binding, it had no effect on binding of this secretagogue, suggesting that a novel cascade not involving a guanine nucleotide-binding protein mediates CCK stimulation of pancreatic secretion.

AB - The molecular events that mediate cholecystokinin (CCK)-stimulated pancreatic secretion are not well defined because of the complex receptor-binding and concentration-response characteristics of this hormone. Functional models of receptor occupancy initiating the cascade leading to secretion have been complicated by the inhibition of secretion effected by supra-maximal concentrations of CCK. Recent report of a CCK analogue that does not exhibit supramaximal inhibition led us to synthesize a similar analogue that could also be radiolabeled for studies of receptor binding and affinity labeling, and for studies of second messenger activity. This probe, D-Tyr-Gly-[(Nle28,31)CCK-26-32]-phenethyl ester, was a fully efficacious secretagogue with no supramaximal inhibition, and, unlike native hormone, bound to a single class of sites present on both acini and membranes. Occupation of this site correlated well with stimulation of secretion. Evidence that this was indeed a CCK-binding site were the abilities of CCK and antagonist L-364,718 to inhibit binding of this analogue. Affinity labeling confirmed the identity of the site mediating secretory stimulation as a M(r) = 85,000-95,000 protein. Whereas the nonhydrolyzable guanosine triphosphate analogue, 5'-guanylyl-imidodiphosphate, was a potent inhibitor of CCK binding, it had no effect on binding of this secretagogue, suggesting that a novel cascade not involving a guanine nucleotide-binding protein mediates CCK stimulation of pancreatic secretion.

UR - http://www.scopus.com/inward/record.url?scp=0024495396&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024495396&partnerID=8YFLogxK

M3 - Article

C2 - 2910915

AN - SCOPUS:0024495396

VL - 83

SP - 321

EP - 325

JO - Journal of Clinical Investigation

JF - Journal of Clinical Investigation

SN - 0021-9738

IS - 1

ER -