Effect of methylglyoxal modification of human α-crystallin on the structure, stability and chaperone function.
α-Crystallin functions as a molecular chaperone and maintains transparency of eye lens by protecting other lens-proteins. Non-enzymatic glycation of α-crystallin by methylglyoxal, plays a crucial role on its chaperone function and structural stability. Our studies showed that methylglyoxal modification even in lower concentration caused significant decrease in chaperone function of α-crystallin as reflected both in thermal aggregation assay and enzyme refolding assay. Thermal denaturation studies showed drastic reduction of denaturation temperature with increase in the degree of modification. Thermodynamic stability studies by urea denaturation assay reflected a decrease of transition midpoint. Quantitatively we found that ΔG° of native α-crystallin decreased from 21.6 kJ/mol to 10.4 kJ/mol due to 72 h modification by 10 mM methylglyoxal. The surface hydrophobicity of α-crystallin after MG modification, was found to be decreased. Circular dichroism spectroscopy revealed conversion of ß-sheet structure to random coil structure. Significant cross-linking was also observed due to methylglyoxal modification of human α-crystallin.
Department of Biochemistry, NRS Medical College and Hospital, Kolkata 700 014, India. s.amukhopadhyay@gmail.com
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D000293:Adolescent; D002648:Child; D005123:Eye / Q000737:chemistry* / Q000378:metabolism; D017127:Glycation End Products, Advanced / Q000737:chemistry* / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D008297:Male; D018832:Molecular Chaperones / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D021122:Protein Subunits / Q000737:chemistry; D011765:Pyruvaldehyde / Q000737:chemistry* / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D038201:alpha-Crystallins / Q000737:chemistry / Q000502:physiology
D017127:Glycation End Products, Advanced; D018832:Molecular Chaperones; D021122:Protein Subunits; D038201:alpha-Crystallins; D011765:Pyruvaldehyde