Structure, antihyperglycemic activity and cellular actions of a novel diglycated human insulin.

F P O'Harte, A C Boyd, A M McKillop, Y H Abdel-Wahab, H McNulty, C R Barnett, J M Conlon, P Højrup, P R Flatt

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Human insulin was glycated under hyperglycemic reducing conditions and a novel diglycated form (M(r) 6135.1 Da) was purified by RP-HPLC. Endoproteinase Glu-C digestion combined with mass spectrometry and automated Edman degradation localized glycation to Gly(1) and Phe(1) of the insulin A- and B-chains, respectively. Intraperitoneal (i.p.) administration of diglycated insulin to mice alone or in combination with glucose (7 nmol/kg) resulted in a 43-61% and 11-34% reduction in glucose lowering activity, respectively, compared with native insulin. Consistent with these findings, diglycated insulin (10(-9) to 10(-7) mol/liter) was 22-38% less effective (P < 0.001) than native insulin in stimulating glucose uptake, glucose oxidation and glycogen production in isolated mouse abdominal muscle.
Original languageEnglish
JournalPeptides
Volume21
Issue number10
Pages (from-to)1519-26
Number of pages7
ISSN0196-9781
DOIs
Publication statusPublished - 1. Oct 2000

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Hypoglycemic Agents
Insulin
Glucose
Abdominal Muscles
Glycogen
Mass spectrometry
Muscle
High Pressure Liquid Chromatography
Degradation
Oxidation

Keywords

  • Amino Acids
  • Animals
  • Biological Transport
  • Blood Glucose
  • Chromatography, High Pressure Liquid
  • Deoxyglucose
  • Glucose
  • Glycogen
  • Glycoproteins
  • Glycosylation
  • Humans
  • Hyperglycemia
  • Injections, Intraperitoneal
  • Insulin
  • Male
  • Mass Spectrometry
  • Mice
  • Mice, Inbred Strains
  • Muscle, Smooth
  • Serine Endopeptidases

Cite this

O'Harte, F. P., Boyd, A. C., McKillop, A. M., Abdel-Wahab, Y. H., McNulty, H., Barnett, C. R., ... Flatt, P. R. (2000). Structure, antihyperglycemic activity and cellular actions of a novel diglycated human insulin. Peptides, 21(10), 1519-26. https://doi.org/10.1016/S0196-9781(00)00306-5
O'Harte, F P ; Boyd, A C ; McKillop, A M ; Abdel-Wahab, Y H ; McNulty, H ; Barnett, C R ; Conlon, J M ; Højrup, P ; Flatt, P R. / Structure, antihyperglycemic activity and cellular actions of a novel diglycated human insulin. In: Peptides. 2000 ; Vol. 21, No. 10. pp. 1519-26.
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O'Harte, FP, Boyd, AC, McKillop, AM, Abdel-Wahab, YH, McNulty, H, Barnett, CR, Conlon, JM, Højrup, P & Flatt, PR 2000, 'Structure, antihyperglycemic activity and cellular actions of a novel diglycated human insulin.', Peptides, vol. 21, no. 10, pp. 1519-26. https://doi.org/10.1016/S0196-9781(00)00306-5

Structure, antihyperglycemic activity and cellular actions of a novel diglycated human insulin. / O'Harte, F P; Boyd, A C; McKillop, A M; Abdel-Wahab, Y H; McNulty, H; Barnett, C R; Conlon, J M; Højrup, P; Flatt, P R.

In: Peptides, Vol. 21, No. 10, 01.10.2000, p. 1519-26.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Structure, antihyperglycemic activity and cellular actions of a novel diglycated human insulin.

AU - O'Harte, F P

AU - Boyd, A C

AU - McKillop, A M

AU - Abdel-Wahab, Y H

AU - McNulty, H

AU - Barnett, C R

AU - Conlon, J M

AU - Højrup, P

AU - Flatt, P R

PY - 2000/10/1

Y1 - 2000/10/1

N2 - Human insulin was glycated under hyperglycemic reducing conditions and a novel diglycated form (M(r) 6135.1 Da) was purified by RP-HPLC. Endoproteinase Glu-C digestion combined with mass spectrometry and automated Edman degradation localized glycation to Gly(1) and Phe(1) of the insulin A- and B-chains, respectively. Intraperitoneal (i.p.) administration of diglycated insulin to mice alone or in combination with glucose (7 nmol/kg) resulted in a 43-61% and 11-34% reduction in glucose lowering activity, respectively, compared with native insulin. Consistent with these findings, diglycated insulin (10(-9) to 10(-7) mol/liter) was 22-38% less effective (P < 0.001) than native insulin in stimulating glucose uptake, glucose oxidation and glycogen production in isolated mouse abdominal muscle.

AB - Human insulin was glycated under hyperglycemic reducing conditions and a novel diglycated form (M(r) 6135.1 Da) was purified by RP-HPLC. Endoproteinase Glu-C digestion combined with mass spectrometry and automated Edman degradation localized glycation to Gly(1) and Phe(1) of the insulin A- and B-chains, respectively. Intraperitoneal (i.p.) administration of diglycated insulin to mice alone or in combination with glucose (7 nmol/kg) resulted in a 43-61% and 11-34% reduction in glucose lowering activity, respectively, compared with native insulin. Consistent with these findings, diglycated insulin (10(-9) to 10(-7) mol/liter) was 22-38% less effective (P < 0.001) than native insulin in stimulating glucose uptake, glucose oxidation and glycogen production in isolated mouse abdominal muscle.

KW - Amino Acids

KW - Animals

KW - Biological Transport

KW - Blood Glucose

KW - Chromatography, High Pressure Liquid

KW - Deoxyglucose

KW - Glucose

KW - Glycogen

KW - Glycoproteins

KW - Glycosylation

KW - Humans

KW - Hyperglycemia

KW - Injections, Intraperitoneal

KW - Insulin

KW - Male

KW - Mass Spectrometry

KW - Mice

KW - Mice, Inbred Strains

KW - Muscle, Smooth

KW - Serine Endopeptidases

U2 - 10.1016/S0196-9781(00)00306-5

DO - 10.1016/S0196-9781(00)00306-5

M3 - Journal article

VL - 21

SP - 1519

EP - 1526

JO - Peptides

JF - Peptides

SN - 0196-9781

IS - 10

ER -

O'Harte FP, Boyd AC, McKillop AM, Abdel-Wahab YH, McNulty H, Barnett CR et al. Structure, antihyperglycemic activity and cellular actions of a novel diglycated human insulin. Peptides. 2000 Oct 1;21(10):1519-26. https://doi.org/10.1016/S0196-9781(00)00306-5