Abstract
In diabetes, protein glycation mostly occurs at intrachain lysine residues resulting in the formation of early stage Amadori products which are finally converted to advance glycation end products (AGEs). Several studies have reported autoantibodies against AGEs in diabetes but not much data are found in respect of Amadori products. In this study, poly-l-lysine (PLL) was glycated with 50 mM glucose and the resultant Amadori products were estimated by fructosamine or nitroblue tetrazolium assay. We report high content of Amadori products in PLL upon glycation. Glycated PLL showed marked hyperchromicity in the UV spectrum, ellipticity changes in CD spectroscopy, and variations in ε-methylene protons shift in NMR. It was better recognized by autoantibodies in type 2 diabetics compared to the native PLL. Induced antibodies against glycated PLL were successfully used to probe early glycation in the IgG isolated from diabetes type 2 patients. Role of Amadori products of glycated proteins in the induction of autoantibodies in type 2 diabetes as well as in associated secondary complications has been discussed.
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Abbreviations
- PLL:
-
Poly-l-lysine
- NBT:
-
Nitroblue tetrazolium
- HMF:
-
5-Hydroxymethylfurfural
- NaBH4 :
-
Sodium borohydride
- AGEs:
-
Advanced glycation end products
- UV:
-
Ultraviolet
- CD spectroscopy:
-
Circular dichroism
- NMR:
-
Nuclear magnetic resonance
- ELISA:
-
Enzyme-linked immunosorbent assay
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Acknowledgments
We are thankful to Sophisticated Analytical Instrument Facility of Central Drug Research Institute, Lucknow (India) for NMR experiments. Thanks are also due to Prof. Jamal Ahmad (Department of Medicine, J.N. Medical College, Aligarh Muslim University) for providing the type 2 diabetes mellitus samples. Financial assistance from UPCST, Lucknow and ICMR, New Delhi to Moinuddin is gratefully acknowledged.
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Ansari, N.A., Moinuddin, Mir, A.R. et al. Role of Early Glycation Amadori Products of Lysine-Rich Proteins in the Production of Autoantibodies in Diabetes Type 2 Patients. Cell Biochem Biophys 70, 857–865 (2014). https://doi.org/10.1007/s12013-014-9991-7
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DOI: https://doi.org/10.1007/s12013-014-9991-7