Fructosylation generates neo-epitopes on human serum albumin.
Hyperglycemia is the defining feature of diabetes mellitus. The persistently high levels of reducing sugars like glucose and fructose cause glycation of various macromolecules in the body. Human serum albumin (HSA), the most abundant serum protein with a myriad of functions, is prone to glycation and consequent alteration in its structural and biological properties. This study aimed to assess the role of fructose-modified human serum albumin as a marker of diabetic pathophysiology. We carried out modification of HSA with fructose and the changes induced were studied by various physicochemical studies. Fructose modified-HSA showed hyperchromicity in UV spectrum and increased AGE-specific fluorescence as well as quenching of tryptophan fluorescence. In SDS-PAGE protein aggregation was seen. Amadori products were detected by NBT. The fructose modified HSA had higher content of carbonyls along with perturbations in secondary structure as revealed by CD and FT-IR. A greater hydrodynamic radius of fructose-modified HSA was evident by DLS measurement. The fructose-modified HSA induced high titre antibodies in experimental animals exhibiting high specificity towards the immunogen.
Department of Biochemistry, J. N. Medical College, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.
Shaziya
S
Allarakha
Department of Biochemistry, J. N. Medical College, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.
Parvez
P
Ahmad
Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.
Mohd
M
Ishtikhar
Department of Medicine, J.N. Medical College, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.
Mohammad Shoaib
MS
Zaheer
Rajeev Gandhi Centre for Diabetes and Endocrinology, J.N. Medical College, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.
Sheelu Shafiq
SS
Siddiqi
Department of Biochemistry, J. N. Medical College, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.
Moinuddin
Department of Biochemistry, J. N. Medical College, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.
Asif
A
Ali
D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D005632:Fructose / Q000378:metabolism*; D006801:Humans; D012709:Serum Albumin / Q000276:immunology*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D000939:Epitopes; D012709:Serum Albumin; D005632:Fructose
AGE; diabetes mellitus; fructose; glycation; human serum albumin