Wagner Mark C, Myslinski Jered, Pratap Shiv, Flores Brittany, Rhodes George, Campos-Bilderback Silvia B, Sandoval Ruben M, Kumar Sudhanshu, Patel Monika, Molitoris Bruce A
Nephrology Division, Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana; and.
The Council of Scientific and Industrial Research Institute of Microbial Technology, Chandigarh, India.
Am J Physiol Renal Physiol. 2016 May 1;310(10):F1089-102. doi: 10.1152/ajprenal.00605.2015. Epub 2016 Feb 17.
Serum albumin is the most abundant plasma protein and has a long half-life due to neonatal Fc receptor (FcRn)-mediated transcytosis by many cell types, including proximal tubule cells of the kidney. Albumin also interacts with, and is modified by, many small and large molecules. Therefore, the focus of the present study was to address the impact of specific known biological albumin modifications on albumin-FcRn binding and cellular handling. Binding at pH 6.0 and 7.4 was performed since FcRn binds albumin strongly at acidic pH and releases it after transcytosis at physiological pH. Equilibrium dissociation constants were measured using microscale thermophoresis. Since studies have shown that glycated albumin is excreted in the urine at a higher rate than unmodified albumin, we studied glucose and methylgloxal modified albumins (21 days). All had reduced affinity to FcRn at pH 6.0, suggesting these albumins would not be returned to the circulation via the transcytotic pathway. To address why modified albumin has reduced affinity, we analyzed the structure of the modified albumins using small-angle X-ray scattering. This analysis showed significant structural changes occurring to albumin with glycation, particularly in the FcRn-binding region, which could explain the reduced affinity to FcRn. These results offer an explanation for enhanced proximal tubule-mediated sorting and clearance of abnormal albumins.
血清白蛋白是最丰富的血浆蛋白,由于新生儿Fc受体(FcRn)介导的多种细胞类型(包括肾近端小管细胞)的转胞吞作用,其半衰期较长。白蛋白还与许多小分子和大分子相互作用并被其修饰。因此,本研究的重点是探讨特定已知生物白蛋白修饰对白蛋白-FcRn结合及细胞处理的影响。由于FcRn在酸性pH下强烈结合白蛋白并在生理pH下转胞吞后释放,因此在pH 6.0和7.4下进行结合实验。使用微量热泳测定平衡解离常数。由于研究表明糖化白蛋白比未修饰的白蛋白以更高的速率从尿液中排出,我们研究了葡萄糖和甲基乙二醛修饰的白蛋白(21天)。所有修饰的白蛋白在pH 6.0时对FcRn的亲和力均降低,这表明这些白蛋白不会通过转胞吞途径返回循环。为了探究修饰后的白蛋白亲和力降低的原因,我们使用小角X射线散射分析了修饰白蛋白的结构。该分析表明,糖化作用使白蛋白发生了显著的结构变化,尤其是在FcRn结合区域,这可以解释其对FcRn亲和力降低的原因。这些结果为近端小管介导的异常白蛋白分选和清除增强提供了解释。