Article

A role for the membrane proteome in human chronic kidney disease erythrocytes

Complutense University of Madrid, Madrid, Madrid, Spain
Translational Research (Impact Factor: 4.04). 07/2012; 160(5):374-83. DOI: 10.1016/j.trsl.2012.06.004
Source: PubMed

ABSTRACT The molecular basis of the reduced half-life of chronic kidney disease (CKD) erythrocytes is unclear. The erythrocyte membrane plays a key role in the erythrocyte mechanical properties and survival. The aim of the present work is to uncover erythrocyte membrane proteins whose expression could be altered in CKD. The erythrocyte membrane subproteome was analyzed by a non-biased approach where the whole set of proteins was simultaneously investigated by 2D fluorescence difference gel electrophoresis without preselection of potential targets. Proteins significantly altered in CKD were identified by mass spectrometry (MS) and results validation was performed by Western blot and confocal microscopy. Nine differentially expressed spots among healthy individuals, non-dialyzed CKD and erythropoietin/dialysis-treated CKD patients were identified by MS/MS corresponding to 5 proteins (beta-adducin, HSP71/72, tropomodulin-1, ezrin, and radixin). Ezrin and radixin were higher in dialyzed CKD patients than in the other 2 groups. Beta-adducin was increased in CKD patients (dialyzed or not). Three spots were normalized in patients on the dialysis/erythropoietin combination compared with non-dialyzed CKD. Among these, a spot corresponding to tropomodulin 1, was found to be of higher abundance in non-dialyzed CKD patients compared with controls or dialyzed CKD. In conclusion, this study identifies changes in erythrocyte membrane proteins in CKD, which may be relevant for the pathogenesis of red cell abnormalities in uremia.

0 Followers
 · 
112 Views
  • [Show abstract] [Hide abstract]
    ABSTRACT: ABSTRACT Chronically haemodialysed end-stage renal disease patients are at high risk of morbidity arising from complications of dialysis, the underlying pathology that has led to renal disease and the complex pathology of chronic kidney disease. Anaemia is commonplace and its origins are multifactorial, involving reduced renal erythropoietin production, accumulation of uremic toxins and an increase in erythrocyte fragility. Oxidative damage is a common risk factor in renal disease and its co-morbidities, and is known to cause erythrocyte fragility. Therefore we have investigated the hypothesis that specific erythrocyte membrane proteins are more oxidised in end-stage renal disease patients and that vitamin C supplementation can ameliorate membrane protein oxidation. Eleven patients and fifteen control subjects were recruited to the study. Patients were supplemented with 2 x 500mg vitamin C per day for four weeks. Erythrocyte membrane proteins were prepared pre- and post-vitamin C supplementation for determination of protein oxidation. Total protein carbonyls were reduced by vitamin C supplementation but not by dialysis when investigated by enzyme linked immunosorbent assay. Using a western blot to detect oxidised proteins, one protein band, later identified as containing ankyrin, was found to be oxidised in patients but not controls and was reduced significantly by 60% in all patients after dialysis and by 20% after vitamin C treatment pre-dialysis. Ankyrin oxidation analysis may be useful in a stratified medicines approach as a possible marker to identify requirements for intervention in dialysis patients.
    Free Radical Research 11/2014; 49(2):1-38. DOI:10.3109/10715762.2014.991725 · 2.99 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: In this minireview, we focus on advances in our knowledge of the human erythrocyte proteome and interactome that have occurred since our seminal review on the topic published in 2007. As will be explained, the number of unique proteins has grown from 751 in 2007 to 2289 as of today. We describe how proteomics and interactomics tools have been used to probe critical protein changes in disorders impacting the blood. The primary example used is the work done on sickle cell disease where biomarkers of severity have been identified, protein changes in the erythrocyte membranes identified, pharmacoproteomic impact of hydroxyurea studied and interactomics used to identify erythrocyte protein changes that are predicted to have the greatest impact on protein interaction networks.
    Experimental Biology and Medicine 05/2013; 238(5):509-18. DOI:10.1177/1535370213488474 · 2.23 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: In humans, when plasma sodium concentration rises slightly beyond 140 mM, vascular endothelium sharply stiffens and nitric oxide release declines. In search of a vascular sodium sensor, the endothelial glycocalyx was identified as being a negatively charged biopolymer capable of selectively buffering sodium ions. Sodium excess damages the glycocalyx and renders vascular endothelium increasingly permeable for sodium. In the long term, sodium accumulates in the interstitium and gradually damages the organism. It was discovered that circulating red blood cells (RBC) 'report' surface properties of the vascular endothelium. To some extent, the RBC glycocalyx mirrors the endothelial glycocalyx. A poor (charge-deprived) endothelial glycocalyx causes a poor RBC glycocalyx and vice versa. This observation led to the assumption that the current state of an individual's vascular endothelium in terms of electrical surface charges and sodium-buffering capabilities could be read simply from a blood sample. Recently, a so-called salt blood test was introduced that quantifies the RBC sodium buffer capacity and thus characterizes the endothelial function. The arguments are outlined in this article spanning a bridge from cellular nano-mechanics to clinical application.
    Nephrology Dialysis Transplantation 12/2013; DOI:10.1093/ndt/gft461 · 3.49 Impact Factor