Ability of GHTD-amide and analogs to enhance insulin activity through zinc chelation and dispersal of insulin oligomers
- Paule, Sarah, Nikolovski, Biljana, Ludeman, Justin, Gray, Robyn, Spiccia, Leone, Zimmet, Paul, Myers, Mark
- Authors: Paule, Sarah , Nikolovski, Biljana , Ludeman, Justin , Gray, Robyn , Spiccia, Leone , Zimmet, Paul , Myers, Mark
- Date: 2009
- Type: Text , Journal article
- Relation: Peptides Vol. 30, no. 6 (2009), p. 1088-1097
- Full Text:
- Reviewed:
- Description: GHTD-amide is a tetrapeptide originally isolated from human urine that has hypoglycemic activity. Insulin occurs in secretory granules of beta cells as zinc-stabilized hexamers and must disperse to monomeric form in order to bind to its receptor. The aim of this study was to identify whether GHTD-amide and an analog called ISF402 (VHTD-amide) reduce blood glucose through enhancement of insulin activity by dispersing oligomers of insulin. Peptides containing the HTD-amide sequence and a free α-amino group were optimal at binding Zn2+ and adopting secondary structure in the presence of Zn2+. Binding was concentration dependent and resulted in a 1:1 Zn:peptide complex. In vitro the tetrapeptides dispersed hexameric insulin to dimers and monomers. GHTD-amide and ISF402 potentiated the activity of hexameric insulin when co-injected into insulin resistant Zucker rats. Injection of peptides with insulin caused reductions in blood glucose and C-peptide significantly larger than achieved with insulin alone, and serum insulin time profiles were also altered consistent with a reduced clearance or enhanced dispersal of the injected insulin. Insulin potentiation by ISF402 was reduced when lispro insulin, which does not form zinc-stabilized hexamers, was used in place of hexameric zinc insulin. In conclusion, GHTD-amide and ISF402 are zinc binding peptides that disperse hexameric insulin in vitro, and potentiate the activity of hexameric insulin more so than monomeric lispro insulin. These results suggest that dispersal of hexameric insulin through chelation of Zn2+ contributes to the hypoglycemic activity of these tetrapeptides. Crown Copyright © 2009.
- Authors: Paule, Sarah , Nikolovski, Biljana , Ludeman, Justin , Gray, Robyn , Spiccia, Leone , Zimmet, Paul , Myers, Mark
- Date: 2009
- Type: Text , Journal article
- Relation: Peptides Vol. 30, no. 6 (2009), p. 1088-1097
- Full Text:
- Reviewed:
- Description: GHTD-amide is a tetrapeptide originally isolated from human urine that has hypoglycemic activity. Insulin occurs in secretory granules of beta cells as zinc-stabilized hexamers and must disperse to monomeric form in order to bind to its receptor. The aim of this study was to identify whether GHTD-amide and an analog called ISF402 (VHTD-amide) reduce blood glucose through enhancement of insulin activity by dispersing oligomers of insulin. Peptides containing the HTD-amide sequence and a free α-amino group were optimal at binding Zn2+ and adopting secondary structure in the presence of Zn2+. Binding was concentration dependent and resulted in a 1:1 Zn:peptide complex. In vitro the tetrapeptides dispersed hexameric insulin to dimers and monomers. GHTD-amide and ISF402 potentiated the activity of hexameric insulin when co-injected into insulin resistant Zucker rats. Injection of peptides with insulin caused reductions in blood glucose and C-peptide significantly larger than achieved with insulin alone, and serum insulin time profiles were also altered consistent with a reduced clearance or enhanced dispersal of the injected insulin. Insulin potentiation by ISF402 was reduced when lispro insulin, which does not form zinc-stabilized hexamers, was used in place of hexameric zinc insulin. In conclusion, GHTD-amide and ISF402 are zinc binding peptides that disperse hexameric insulin in vitro, and potentiate the activity of hexameric insulin more so than monomeric lispro insulin. These results suggest that dispersal of hexameric insulin through chelation of Zn2+ contributes to the hypoglycemic activity of these tetrapeptides. Crown Copyright © 2009.
GHTD-amide : A naturally occurring beta cell-derived peptide with hypoglycemic activity
- Paule, Sarah, Nikolovski, Biljana, Gray, Robyn, Ludeman, Justin, Freemantle, A., Spark, R. A., Kerr, J. B., Ng, F. M., Zimmet, Paul, Myers, Mark
- Authors: Paule, Sarah , Nikolovski, Biljana , Gray, Robyn , Ludeman, Justin , Freemantle, A. , Spark, R. A. , Kerr, J. B. , Ng, F. M. , Zimmet, Paul , Myers, Mark
- Date: 2009
- Type: Text , Journal article
- Relation: Peptides Vol. 30, no. 5 (2009), p. 955-961
- Full Text:
- Reviewed:
- Description: in the early 1970s, a peptide fraction with insulin potentiating activity was purified from human urine but the identity and origins of the active constituent remained unknown. Here we identify the active component and characterize its origins. The active peptide was identified as an alpha amidated tetrapeptide with the sequence GHTD-amide. The peptide was synthesized and tested for stimulation of glycogen synthesis and insulin potentiation by insulin tolerance testing in insulin-deficient rats, which confirmed GHTD-amide as the active peptide. Tissue localization using a peptide-specific anti-serum and epifluorescent and confocal microscopy showed decoration of pancreatic islets but not other tissues. Confocal microscopy revealed co-localization with insulin and immunogold and electron microscopy showed localization to dense core secretory granules. Consistent with these observations GHTD-amide was found in media conditioned by MIN6 islet beta cells. Sequence database searching found no annotated protein in the human proteome encoding a potential precursor for GHTD-amide. We conclude that the insulin potentiating activity originally described in human urine is attributable to the tetrapeptide GHTD-amide. GHTD-amide is a novel peptide produced by pancreatic beta cells and no precursor protein is present in the annotated human proteome. Stimulation of glycogen synthesis and co-localization with insulin in beta cells suggest that GHTD-amide may play a role in glucose homeostasis by enhancing insulin action and glucose storage in tissues. (C) 2008 Elsevier Inc. All rights reserved.
- Authors: Paule, Sarah , Nikolovski, Biljana , Gray, Robyn , Ludeman, Justin , Freemantle, A. , Spark, R. A. , Kerr, J. B. , Ng, F. M. , Zimmet, Paul , Myers, Mark
- Date: 2009
- Type: Text , Journal article
- Relation: Peptides Vol. 30, no. 5 (2009), p. 955-961
- Full Text:
- Reviewed:
- Description: in the early 1970s, a peptide fraction with insulin potentiating activity was purified from human urine but the identity and origins of the active constituent remained unknown. Here we identify the active component and characterize its origins. The active peptide was identified as an alpha amidated tetrapeptide with the sequence GHTD-amide. The peptide was synthesized and tested for stimulation of glycogen synthesis and insulin potentiation by insulin tolerance testing in insulin-deficient rats, which confirmed GHTD-amide as the active peptide. Tissue localization using a peptide-specific anti-serum and epifluorescent and confocal microscopy showed decoration of pancreatic islets but not other tissues. Confocal microscopy revealed co-localization with insulin and immunogold and electron microscopy showed localization to dense core secretory granules. Consistent with these observations GHTD-amide was found in media conditioned by MIN6 islet beta cells. Sequence database searching found no annotated protein in the human proteome encoding a potential precursor for GHTD-amide. We conclude that the insulin potentiating activity originally described in human urine is attributable to the tetrapeptide GHTD-amide. GHTD-amide is a novel peptide produced by pancreatic beta cells and no precursor protein is present in the annotated human proteome. Stimulation of glycogen synthesis and co-localization with insulin in beta cells suggest that GHTD-amide may play a role in glucose homeostasis by enhancing insulin action and glucose storage in tissues. (C) 2008 Elsevier Inc. All rights reserved.
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