Ribosylation of bovine serum albumin induces ROS accumulation and cell death in cancer line (MCF-7)
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ORIGINAL PAPER
Ribosylation of bovine serum albumin induces ROS accumulation and cell death in cancer line (MCF-7) Mohd Shahnawaz Khan • Sourabh Dwivedi • Medha Priyadarshini • Shams Tabrez • Maqsood Ahmed Siddiqui • Haseeb Jagirdar • Abdulrahman M. Al-Senaidy • Abdulaziz A. Al-Khedhairy • Javed Musarrat
Received: 28 April 2013 / Revised: 30 July 2013 / Accepted: 10 September 2013 / Published online: 12 November 2013 Ó European Biophysical Societies’ Association 2013
Abstract Formation of advanced glycation end products (AGE) is crucially involved in the several pathophysiologies associated with ageing and diabetes, for example arthritis, atherosclerosis, chronic renal insufficiency, Alzheimer’s disease, nephropathy, neuropathy, and cataracts. Because of devastating effects of AGE and the significance of bovine serum albumin (BSA) as a transport protein, this study was designed to investigate glycation-induced structural modifications in BSA and their functional consequences in breast cancer cell line (MCF-7). We incubated D-ribose with BSA and monitored formation of D-ribose-glycated BSA by observing changes in the intensity of fluorescence at 410 nm. NBT (nitro blue tetrazolium) assay was performed to confirm formation of keto-amine during glycation. Absorbance at 540 nm (fructosamine) increased markedly with time. Furthermore, intrinsic protein and 8-anilino-1-naphthalenesulfonate (ANS) fluorescence revealed marked
conformational changes in BSA upon ribosylation. In addition, a fluorescence assay with thioflavin T (ThT) revealed a remarkable increase in fluorescence at 485 nm in the presence of glycated BSA. This suggests that glycation with Dribose induced aggregation of BSA into amyloid-like deposits. Circular dichroism (CD) study of native and ribosylated BSA revealed molten globule formation in the glycation pathway of BSA. Functional consequences of ribosylated BSA on cancer cell line, MCF-7 was studied by MTT assay and ROS estimation. The results revealed cytotoxicity of ribosylated BSA on MCF-7 cells.
M. S. Khan (&) H. Jagirdar A. M. Al-Senaidy Department of Biochemistry, College of Science, King Saud University, Riyadh, Kingdom of Saudi Arabia e-mail: [email protected]
Non-enzymatic glycation is a complex cascade of reactions initiated by condensation of reducing sugars with the free amino groups of proteins to form reversible Schiff’s bases which undergo rearrangement to form relatively stable Amadori products. Amadori products, over a period of time, undergo a series of reactions involving multiple dehydration, fragmentation, and oxidative modification via highly reactive dicarbonyl intermediates to form stable, heterogeneous adducts called advanced glycation end products (AGE) (Sing et al. 2001; Baynes et al. 1989; Monnier 1989). Although AGE formation occurs during the normal ageing process, it is accelerated under hyperglycemic conditions. Further, it has been shown that formation of AGE in vivo contributes to several pathophysiologies associated with ageing and diabetes mellitus, for examp
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