Identification of a bioactive domain in the amino-terminus of glucose-dependent insulinotropic polypeptide (GIP)

Simon A. Hinke, Susanne Manhart, Nathalie Pamir, Hans Ulrich Demuth, Richard W. Gelling, Raymond A. Pederson, Christopher H S McIntosh

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

The incretins are a class of hormones released from the small bowel that act on the endocrine pancreas to potentiate insulin secretion in a glucose-dependent manner. Due to the requirement for an elevated glucose concentration for activity, the incretins, glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1, have potential in the treatment of non-insulin-dependent diabetes mellitus. A series of synthetic peptide GIP fragments was generated for the purpose of elucidating the bioactive domain of the molecule. Peptides were screened for stimulation of cyclic AMP (cAMP) accumulation in Chinese hamster ovary cells transfected with the rat islet GIP receptor. Of the GIP fragments tested, GIP1-14 and GIP19-30 demonstrated the greatest cAMP-stimulating ability over the range of concentrations tested (up to 20 μM). In contrast, GIP fragments corresponding to amino acids 15-42, 15-30, 16-30 and 17-30 all demonstrated weak antagonism of GIP1-42 activity. Competitive-binding displacement studies indicated that these peptides were low-affinity ligands for the GIP receptor. To examine biological activity in vivo, a bioassay was developed in the anesthetized rat. Intravenous infusion of GIP1-42 (1 pmol/min/100 g) with a concurrent intraperitoneal glucose load (1 g/kg) significantly reduced circulating blood glucose excursions through stimulation of insulin release. Higher doses of GIP1-14 and GIP19-30 (100 pmol/min/100 g) also reduced blood glucose excursions.

Original languageEnglish (US)
Pages (from-to)143-155
Number of pages13
JournalBiochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
Volume1547
Issue number1
DOIs
StatePublished - May 5 2001
Externally publishedYes

Fingerprint

Glucose
Peptides
Incretins
Cyclic AMP
Blood Glucose
Rats
Gastric Inhibitory Polypeptide
Insulin
Glucagon-Like Peptide 1
Competitive Binding
Bioassay
Polypeptides
Medical problems
Cricetulus
Bioactivity
Islets of Langerhans
Intravenous Infusions
Biological Assay
Type 2 Diabetes Mellitus
Ovary

Keywords

  • Adenylyl cyclase
  • Bioassay
  • Competitive binding
  • Cyclic adenosine monophosphate
  • Enteroinsular axis
  • Perfused pancreas
  • Structure-activity relationship

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Structural Biology
  • Biophysics

Cite this

Identification of a bioactive domain in the amino-terminus of glucose-dependent insulinotropic polypeptide (GIP). / Hinke, Simon A.; Manhart, Susanne; Pamir, Nathalie; Demuth, Hans Ulrich; W. Gelling, Richard; Pederson, Raymond A.; McIntosh, Christopher H S.

In: Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology, Vol. 1547, No. 1, 05.05.2001, p. 143-155.

Research output: Contribution to journalArticle

Hinke, Simon A. ; Manhart, Susanne ; Pamir, Nathalie ; Demuth, Hans Ulrich ; W. Gelling, Richard ; Pederson, Raymond A. ; McIntosh, Christopher H S. / Identification of a bioactive domain in the amino-terminus of glucose-dependent insulinotropic polypeptide (GIP). In: Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology. 2001 ; Vol. 1547, No. 1. pp. 143-155.
@article{bcdb2533c92d4476b6a802ffb6c25d9e,
title = "Identification of a bioactive domain in the amino-terminus of glucose-dependent insulinotropic polypeptide (GIP)",
abstract = "The incretins are a class of hormones released from the small bowel that act on the endocrine pancreas to potentiate insulin secretion in a glucose-dependent manner. Due to the requirement for an elevated glucose concentration for activity, the incretins, glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1, have potential in the treatment of non-insulin-dependent diabetes mellitus. A series of synthetic peptide GIP fragments was generated for the purpose of elucidating the bioactive domain of the molecule. Peptides were screened for stimulation of cyclic AMP (cAMP) accumulation in Chinese hamster ovary cells transfected with the rat islet GIP receptor. Of the GIP fragments tested, GIP1-14 and GIP19-30 demonstrated the greatest cAMP-stimulating ability over the range of concentrations tested (up to 20 μM). In contrast, GIP fragments corresponding to amino acids 15-42, 15-30, 16-30 and 17-30 all demonstrated weak antagonism of GIP1-42 activity. Competitive-binding displacement studies indicated that these peptides were low-affinity ligands for the GIP receptor. To examine biological activity in vivo, a bioassay was developed in the anesthetized rat. Intravenous infusion of GIP1-42 (1 pmol/min/100 g) with a concurrent intraperitoneal glucose load (1 g/kg) significantly reduced circulating blood glucose excursions through stimulation of insulin release. Higher doses of GIP1-14 and GIP19-30 (100 pmol/min/100 g) also reduced blood glucose excursions.",
keywords = "Adenylyl cyclase, Bioassay, Competitive binding, Cyclic adenosine monophosphate, Enteroinsular axis, Perfused pancreas, Structure-activity relationship",
author = "Hinke, {Simon A.} and Susanne Manhart and Nathalie Pamir and Demuth, {Hans Ulrich} and {W. Gelling}, Richard and Pederson, {Raymond A.} and McIntosh, {Christopher H S}",
year = "2001",
month = "5",
day = "5",
doi = "10.1016/S0167-4838(01)00181-9",
language = "English (US)",
volume = "1547",
pages = "143--155",
journal = "Biochimica et Biophysica Acta - Proteins and Proteomics",
issn = "1570-9639",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Identification of a bioactive domain in the amino-terminus of glucose-dependent insulinotropic polypeptide (GIP)

AU - Hinke, Simon A.

AU - Manhart, Susanne

AU - Pamir, Nathalie

AU - Demuth, Hans Ulrich

AU - W. Gelling, Richard

AU - Pederson, Raymond A.

AU - McIntosh, Christopher H S

PY - 2001/5/5

Y1 - 2001/5/5

N2 - The incretins are a class of hormones released from the small bowel that act on the endocrine pancreas to potentiate insulin secretion in a glucose-dependent manner. Due to the requirement for an elevated glucose concentration for activity, the incretins, glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1, have potential in the treatment of non-insulin-dependent diabetes mellitus. A series of synthetic peptide GIP fragments was generated for the purpose of elucidating the bioactive domain of the molecule. Peptides were screened for stimulation of cyclic AMP (cAMP) accumulation in Chinese hamster ovary cells transfected with the rat islet GIP receptor. Of the GIP fragments tested, GIP1-14 and GIP19-30 demonstrated the greatest cAMP-stimulating ability over the range of concentrations tested (up to 20 μM). In contrast, GIP fragments corresponding to amino acids 15-42, 15-30, 16-30 and 17-30 all demonstrated weak antagonism of GIP1-42 activity. Competitive-binding displacement studies indicated that these peptides were low-affinity ligands for the GIP receptor. To examine biological activity in vivo, a bioassay was developed in the anesthetized rat. Intravenous infusion of GIP1-42 (1 pmol/min/100 g) with a concurrent intraperitoneal glucose load (1 g/kg) significantly reduced circulating blood glucose excursions through stimulation of insulin release. Higher doses of GIP1-14 and GIP19-30 (100 pmol/min/100 g) also reduced blood glucose excursions.

AB - The incretins are a class of hormones released from the small bowel that act on the endocrine pancreas to potentiate insulin secretion in a glucose-dependent manner. Due to the requirement for an elevated glucose concentration for activity, the incretins, glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1, have potential in the treatment of non-insulin-dependent diabetes mellitus. A series of synthetic peptide GIP fragments was generated for the purpose of elucidating the bioactive domain of the molecule. Peptides were screened for stimulation of cyclic AMP (cAMP) accumulation in Chinese hamster ovary cells transfected with the rat islet GIP receptor. Of the GIP fragments tested, GIP1-14 and GIP19-30 demonstrated the greatest cAMP-stimulating ability over the range of concentrations tested (up to 20 μM). In contrast, GIP fragments corresponding to amino acids 15-42, 15-30, 16-30 and 17-30 all demonstrated weak antagonism of GIP1-42 activity. Competitive-binding displacement studies indicated that these peptides were low-affinity ligands for the GIP receptor. To examine biological activity in vivo, a bioassay was developed in the anesthetized rat. Intravenous infusion of GIP1-42 (1 pmol/min/100 g) with a concurrent intraperitoneal glucose load (1 g/kg) significantly reduced circulating blood glucose excursions through stimulation of insulin release. Higher doses of GIP1-14 and GIP19-30 (100 pmol/min/100 g) also reduced blood glucose excursions.

KW - Adenylyl cyclase

KW - Bioassay

KW - Competitive binding

KW - Cyclic adenosine monophosphate

KW - Enteroinsular axis

KW - Perfused pancreas

KW - Structure-activity relationship

UR - http://www.scopus.com/inward/record.url?scp=0035810678&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035810678&partnerID=8YFLogxK

U2 - 10.1016/S0167-4838(01)00181-9

DO - 10.1016/S0167-4838(01)00181-9

M3 - Article

C2 - 11343800

AN - SCOPUS:0035810678

VL - 1547

SP - 143

EP - 155

JO - Biochimica et Biophysica Acta - Proteins and Proteomics

JF - Biochimica et Biophysica Acta - Proteins and Proteomics

SN - 1570-9639

IS - 1

ER -