Effects of in vitro glycation on Fe3+ binding and Fe3+ isoforms of transferrin

Clinical Chemistry
Ann Van CampenhoutBegoña Manuel-Y-Keenoy

Abstract

In diabetes, protein function is altered by glycation, but the impact on the Fe3+ binding and antioxidant functions of transferrin (Tf) is largely unknown. The aim of the present study was to investigate the effects of glycation on the distribution of Fe3+ on the two Fe3+ -binding sites of Tf. In vitro glycation of Tf was accomplished by preincubation with glucose for 14 days. Tf was loaded with Fe3+ compounds to achieve theoretical Tf Fe3+ saturations of 32%, 64%, and 96% (monitored by spectrophotometry). Fe3+ -Tf isoforms were separated by isoelectric focusing. Fe3+ binding was highest when Tf was incubated with Fe:nitrilotriacetic acid and reached a steady state overnight. Increasing the Fe3+ load led to a shift of isoform profile toward the diferric form (Fe2-Tf): in freshly prepared Tf, Fe2-Tf represented 6%, 30%, and 66% of all isoforms at 32%, 64%, and 96% theoretical Fe3+ saturation, respectively. Fe3+ was equally distributed to the monoferric Tf forms with Fe3+ bound to the amino (Fe1N-Tf) and carboxy termini (Fe1C-Tf). Glycation decreased binding of Fe3+ to Tf (monitored at 450 nm). At low theoretical Fe3+ saturation (32%), glycation increased the mean (SD) proportion of Fe2-Tf: 18 (3)% in the presence of 33.3 mmol/L ...Continue Reading

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Citations

May 22, 2013·Alcohol and Alcoholism : International Journal of the Medical Council on Alcoholism·Anders Helander, Naama Kenan Modén
Dec 15, 2010·Biotechnology Advances·Martin E BrandsmaShengwu Ma
Mar 4, 2006·Clinica Chimica Acta; International Journal of Clinical Chemistry·Ann Van CampenhoutBegoña Manuel-y-Keenoy
May 9, 2006·Free Radical Biology & Medicine·Ann Van CampenhoutBegoña Manuel-y-Keenoy
Jan 5, 2014·PLoS Pathogens·J Christian Pérez, Alexander D Johnson
Jun 20, 2014·PLoS Pathogens·Halie K MillerVictoria Auerbuch
Feb 15, 2005·Drug Discovery Today·Peter T GommeJoseph Bertolini
Oct 12, 2019·Metallomics : Integrated Biometal Science·Nathaniel DziubaPaul A Lindahl
Jan 29, 2011·Metallomics : Integrated Biometal Science·Mechthild GrebeAndreas Prange
Nov 2, 2011·Metallomics : Integrated Biometal Science·Yoana Nuevo-OrdoñezA Sanz-Medel
Jun 18, 2021·Diabetes, Metabolic Syndrome and Obesity : Targets and Therapy·Yanqi MaSongbo Fu

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