Abstract
Increased glucose concentration in diabetes mellitus causes glycation of several proteins, leading to changes in their properties. Although glycation-induced functional modification of myoglobin is known, structural modification of the protein has not yet been reported. Here, we have studied glucose-modified structural changes of the heme protein. After in vitro glycation of metmyoglobin (Mb) by glucose at 25°C for 6 days, glycated myoglobin (GMb) and unchanged Mb have been separated by ion exchange (BioRex 70) chromatography, and their properties have been compared. Compared to Mb, GMb exhibits increased absorbance around 280 nm and enhanced fluorescence emission with excitation at 285 nm. Fluorescence quenching experiments of the proteins by acrylamide and KI indicate that more surface accessible tryptophan residues are exposed in GMb. CD spectroscopic study reveals a change in the secondary structure of GMb with decreased α-helix content. 1-anilino-naphthaline-8-sulfonate (ANS) binding with Mb and GMb indicates that glycation increases hydrophobicity of the heme protein. GMb appears to be less stable with respect to thermal denaturation and differential calorimetry experiments. Heme-globin linkage becomes weaker in GMb, as shown by spectroscopic and gel electrophoresis experiments. A correlation between glycation-induced structural and functional modifications of the heme protein has been suggested.
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Acknowledgments
R.S gets a Fellowship from the UGC-UPE scheme of the University of Calcutta. A part of the study was supported by a grant from the Council of Scientific and Industrial Research, New Delhi [grant No. 38(1129)/03/EMR-II]. Thanks are due to Dr. N.S. Chatterjee, National Institute of Cholera and Enteric Diseases, Kolkata for allowing us to use Differential Scanning Calorimeter N-DSCII (Calorimetric Scientific Corporation) and Prof. U. Chaudhuri, Department of Biophysics, Molecular Biology & Bioinformatics, C.U for helpful discussions.
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Roy, A., Sil, R. & Chakraborti, A.S. Non-enzymatic glycation induces structural modifications of myoglobin. Mol Cell Biochem 338, 105–114 (2010). https://doi.org/10.1007/s11010-009-0343-7
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DOI: https://doi.org/10.1007/s11010-009-0343-7