Identification of amadori-modified plasma proteins in type 2 diabetes and the effect of short-term intensive insulin treatment

Abdul Jaleel, Panagiotis Halvatsiotis, Brian Willlamson, Peter Juhasz, Stephen Martin, K Sreekumaran Nair

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

OBJECTIVE - Growing evidence supports that nonenzymatic glycation products may cause hyperglycemia-induced diabetes complications. Amadori-modified proteins are the intermediate products of nonenzymatic glycation and constitute the forms of glycated proteins in diabetes. The objective of the current study was to utilize two-dimensional gel electrophoresis, Western blot, and mass spectrometry to identify Amadori-modified plasma proteins in type 2 diabetic patients with poor glycemic control and assess the impact of short-term insulin treatment on the glycation of these proteins. RESEARCH DESIGN AND METHODS - We compared eight type 2 diabetic subjects (aged 56 ± 3 years and BMI 29.7 ± 0.9 kg/m 2) with an average diabetes duration of 8.5 years (range 3-19) with equal numbers of weight-matched (aged 56 ± 2 years and BMI 30.1 ± 10.0 kg/m 2) and lean (aged 58 ± 2 years and BMI 25 ± 00.5 kg/m 2) nondiabetic subjects who have no first-degree relatives with diabetes. Two separate blood samples were collected from the type 2 diabetic subjects, one following 2 weeks of withdrawal of all antidiabetic medications (T 2D-; plasma glucose 12.6 ± 1.0 mmol/l) and another following 10 days of intensive insulin treatment (T 2D+; plasma glucose 5.5 ± 0.2 mmol/l). Plasma proteins were separated using single and two-dimensional gel electrophoresis. Western blot analysis was performed, and several proteins, which reacted with the Amadori-antibody (1-deoxyfructosyl lysine), were identified by tandem mass spectrometry. RESULTS - No significant differences in the glycation of proteins between the obese and lean groups were noted, but type 2 diabetic patients had several proteins with higher glycation than the control groups. We identified 12 plasma proteins with reduced reaction to the anti-Amadori antibody upon intensive insulin treatment. A significant (P < 0.03) difference in Amadori modification was observed between the T 2D- and control subjects for all these proteins except the Ig light chain. Insulin treatment reduced Amadori modification of albumin (23.2%, P < 0.02), fibrin (34.6%, P < 0,001), Ig heavy chain constant region (20.7%, P < 0.05), transferrin (25.4%, P < 0.04), and Ig light chain (13%, P < 0.02). In addition, Western blot analysis of two-dimensional gel electrophoresis identified α-fibrinogen precursor, β-fibrinogen precursor, fibrinogen γβ chain precursor, hemopexin, vitamin D binding protein, and serine protease inhibitor as proteins with a reduced reaction to anti-Amadori antibody upon intensive insulin treatment. CONCLUSIONS - The current approach offers the opportunity to identify Amadori modification of many proteins that may cause functional alterations and offers the potential for monitoring short-term glycemic control in diabetic patients.

Original languageEnglish (US)
Pages (from-to)645-652
Number of pages8
JournalDiabetes Care
Volume28
Issue number3
DOIs
StatePublished - Mar 2005

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Type 2 Diabetes Mellitus
Blood Proteins
Insulin
Electrophoresis, Gel, Two-Dimensional
Proteins
Fibrinogen
Western Blotting
Therapeutics
Anti-Idiotypic Antibodies
Hemopexin
Vitamin D-Binding Protein
Light
Glucose
Immunoglobulin Heavy Chains
Serine Proteinase Inhibitors
Diabetes Complications
Transferrin
Tandem Mass Spectrometry
Leptin
Fibrin

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism

Cite this

Identification of amadori-modified plasma proteins in type 2 diabetes and the effect of short-term intensive insulin treatment. / Jaleel, Abdul; Halvatsiotis, Panagiotis; Willlamson, Brian; Juhasz, Peter; Martin, Stephen; Nair, K Sreekumaran.

In: Diabetes Care, Vol. 28, No. 3, 03.2005, p. 645-652.

Research output: Contribution to journalArticle

Jaleel, Abdul ; Halvatsiotis, Panagiotis ; Willlamson, Brian ; Juhasz, Peter ; Martin, Stephen ; Nair, K Sreekumaran. / Identification of amadori-modified plasma proteins in type 2 diabetes and the effect of short-term intensive insulin treatment. In: Diabetes Care. 2005 ; Vol. 28, No. 3. pp. 645-652.
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abstract = "OBJECTIVE - Growing evidence supports that nonenzymatic glycation products may cause hyperglycemia-induced diabetes complications. Amadori-modified proteins are the intermediate products of nonenzymatic glycation and constitute the forms of glycated proteins in diabetes. The objective of the current study was to utilize two-dimensional gel electrophoresis, Western blot, and mass spectrometry to identify Amadori-modified plasma proteins in type 2 diabetic patients with poor glycemic control and assess the impact of short-term insulin treatment on the glycation of these proteins. RESEARCH DESIGN AND METHODS - We compared eight type 2 diabetic subjects (aged 56 ± 3 years and BMI 29.7 ± 0.9 kg/m 2) with an average diabetes duration of 8.5 years (range 3-19) with equal numbers of weight-matched (aged 56 ± 2 years and BMI 30.1 ± 10.0 kg/m 2) and lean (aged 58 ± 2 years and BMI 25 ± 00.5 kg/m 2) nondiabetic subjects who have no first-degree relatives with diabetes. Two separate blood samples were collected from the type 2 diabetic subjects, one following 2 weeks of withdrawal of all antidiabetic medications (T 2D-; plasma glucose 12.6 ± 1.0 mmol/l) and another following 10 days of intensive insulin treatment (T 2D+; plasma glucose 5.5 ± 0.2 mmol/l). Plasma proteins were separated using single and two-dimensional gel electrophoresis. Western blot analysis was performed, and several proteins, which reacted with the Amadori-antibody (1-deoxyfructosyl lysine), were identified by tandem mass spectrometry. RESULTS - No significant differences in the glycation of proteins between the obese and lean groups were noted, but type 2 diabetic patients had several proteins with higher glycation than the control groups. We identified 12 plasma proteins with reduced reaction to the anti-Amadori antibody upon intensive insulin treatment. A significant (P < 0.03) difference in Amadori modification was observed between the T 2D- and control subjects for all these proteins except the Ig light chain. Insulin treatment reduced Amadori modification of albumin (23.2{\%}, P < 0.02), fibrin (34.6{\%}, P < 0,001), Ig heavy chain constant region (20.7{\%}, P < 0.05), transferrin (25.4{\%}, P < 0.04), and Ig light chain (13{\%}, P < 0.02). In addition, Western blot analysis of two-dimensional gel electrophoresis identified α-fibrinogen precursor, β-fibrinogen precursor, fibrinogen γβ chain precursor, hemopexin, vitamin D binding protein, and serine protease inhibitor as proteins with a reduced reaction to anti-Amadori antibody upon intensive insulin treatment. CONCLUSIONS - The current approach offers the opportunity to identify Amadori modification of many proteins that may cause functional alterations and offers the potential for monitoring short-term glycemic control in diabetic patients.",
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T1 - Identification of amadori-modified plasma proteins in type 2 diabetes and the effect of short-term intensive insulin treatment

AU - Jaleel, Abdul

AU - Halvatsiotis, Panagiotis

AU - Willlamson, Brian

AU - Juhasz, Peter

AU - Martin, Stephen

AU - Nair, K Sreekumaran

PY - 2005/3

Y1 - 2005/3

N2 - OBJECTIVE - Growing evidence supports that nonenzymatic glycation products may cause hyperglycemia-induced diabetes complications. Amadori-modified proteins are the intermediate products of nonenzymatic glycation and constitute the forms of glycated proteins in diabetes. The objective of the current study was to utilize two-dimensional gel electrophoresis, Western blot, and mass spectrometry to identify Amadori-modified plasma proteins in type 2 diabetic patients with poor glycemic control and assess the impact of short-term insulin treatment on the glycation of these proteins. RESEARCH DESIGN AND METHODS - We compared eight type 2 diabetic subjects (aged 56 ± 3 years and BMI 29.7 ± 0.9 kg/m 2) with an average diabetes duration of 8.5 years (range 3-19) with equal numbers of weight-matched (aged 56 ± 2 years and BMI 30.1 ± 10.0 kg/m 2) and lean (aged 58 ± 2 years and BMI 25 ± 00.5 kg/m 2) nondiabetic subjects who have no first-degree relatives with diabetes. Two separate blood samples were collected from the type 2 diabetic subjects, one following 2 weeks of withdrawal of all antidiabetic medications (T 2D-; plasma glucose 12.6 ± 1.0 mmol/l) and another following 10 days of intensive insulin treatment (T 2D+; plasma glucose 5.5 ± 0.2 mmol/l). Plasma proteins were separated using single and two-dimensional gel electrophoresis. Western blot analysis was performed, and several proteins, which reacted with the Amadori-antibody (1-deoxyfructosyl lysine), were identified by tandem mass spectrometry. RESULTS - No significant differences in the glycation of proteins between the obese and lean groups were noted, but type 2 diabetic patients had several proteins with higher glycation than the control groups. We identified 12 plasma proteins with reduced reaction to the anti-Amadori antibody upon intensive insulin treatment. A significant (P < 0.03) difference in Amadori modification was observed between the T 2D- and control subjects for all these proteins except the Ig light chain. Insulin treatment reduced Amadori modification of albumin (23.2%, P < 0.02), fibrin (34.6%, P < 0,001), Ig heavy chain constant region (20.7%, P < 0.05), transferrin (25.4%, P < 0.04), and Ig light chain (13%, P < 0.02). In addition, Western blot analysis of two-dimensional gel electrophoresis identified α-fibrinogen precursor, β-fibrinogen precursor, fibrinogen γβ chain precursor, hemopexin, vitamin D binding protein, and serine protease inhibitor as proteins with a reduced reaction to anti-Amadori antibody upon intensive insulin treatment. CONCLUSIONS - The current approach offers the opportunity to identify Amadori modification of many proteins that may cause functional alterations and offers the potential for monitoring short-term glycemic control in diabetic patients.

AB - OBJECTIVE - Growing evidence supports that nonenzymatic glycation products may cause hyperglycemia-induced diabetes complications. Amadori-modified proteins are the intermediate products of nonenzymatic glycation and constitute the forms of glycated proteins in diabetes. The objective of the current study was to utilize two-dimensional gel electrophoresis, Western blot, and mass spectrometry to identify Amadori-modified plasma proteins in type 2 diabetic patients with poor glycemic control and assess the impact of short-term insulin treatment on the glycation of these proteins. RESEARCH DESIGN AND METHODS - We compared eight type 2 diabetic subjects (aged 56 ± 3 years and BMI 29.7 ± 0.9 kg/m 2) with an average diabetes duration of 8.5 years (range 3-19) with equal numbers of weight-matched (aged 56 ± 2 years and BMI 30.1 ± 10.0 kg/m 2) and lean (aged 58 ± 2 years and BMI 25 ± 00.5 kg/m 2) nondiabetic subjects who have no first-degree relatives with diabetes. Two separate blood samples were collected from the type 2 diabetic subjects, one following 2 weeks of withdrawal of all antidiabetic medications (T 2D-; plasma glucose 12.6 ± 1.0 mmol/l) and another following 10 days of intensive insulin treatment (T 2D+; plasma glucose 5.5 ± 0.2 mmol/l). Plasma proteins were separated using single and two-dimensional gel electrophoresis. Western blot analysis was performed, and several proteins, which reacted with the Amadori-antibody (1-deoxyfructosyl lysine), were identified by tandem mass spectrometry. RESULTS - No significant differences in the glycation of proteins between the obese and lean groups were noted, but type 2 diabetic patients had several proteins with higher glycation than the control groups. We identified 12 plasma proteins with reduced reaction to the anti-Amadori antibody upon intensive insulin treatment. A significant (P < 0.03) difference in Amadori modification was observed between the T 2D- and control subjects for all these proteins except the Ig light chain. Insulin treatment reduced Amadori modification of albumin (23.2%, P < 0.02), fibrin (34.6%, P < 0,001), Ig heavy chain constant region (20.7%, P < 0.05), transferrin (25.4%, P < 0.04), and Ig light chain (13%, P < 0.02). In addition, Western blot analysis of two-dimensional gel electrophoresis identified α-fibrinogen precursor, β-fibrinogen precursor, fibrinogen γβ chain precursor, hemopexin, vitamin D binding protein, and serine protease inhibitor as proteins with a reduced reaction to anti-Amadori antibody upon intensive insulin treatment. CONCLUSIONS - The current approach offers the opportunity to identify Amadori modification of many proteins that may cause functional alterations and offers the potential for monitoring short-term glycemic control in diabetic patients.

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