Ribosylation of bovine serum albumin induces ROS accumulation and cell death in cancer line (MCF-7).
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 D-ribose 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.
Department of Biochemistry, College of Science, King Saud University, Riyadh, Kingdom of Saudi Arabia, moskhan@ksu.edu.sa.
Mohd Shahnawaz
MS
Khan
Sourabh
S
Dwivedi
Medha
M
Priyadarshini
Shams
S
Tabrez
Maqsood Ahmed
MA
Siddiqui
Haseeb
H
Jagirdar
Abdulrahman M
AM
Al-Senaidy
Abdulaziz A
AA
Al-Khedhairy
Javed
J
Musarrat
D000818:Animals; D002417:Cattle; D016923:Cell Death / Q000187:drug effects; D006031:Glycosylation; D061986:MCF-7 Cells; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012266:Ribose / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism* / Q000494:pharmacology*
D017382:Reactive Oxygen Species; D012710:Serum Albumin, Bovine; D012266:Ribose