Characterization of the glycated human cerebrospinal fluid proteome.
Protein glycation is a nonenzymatic modification that involves pathological functions in neurological diseases. Despite the high number of studies showing accumulation of advanced end glycation products (AGEs) at clinical stage, there is a lack of knowledge about which proteins are modified, where those modifications occur, and to what extent. The goal of this study was to achieve a comprehensive characterization of proteins modified by early glycation in human cerebrospinal fluid (CSF). Approaches based on glucose diferential labeling and mass spectrometry have been applied to evaluate the glycated CSF proteome at two physiological conditions: native glucose level and in vitro high glucose content. For both purposes, detection of glycated proteins was carried out by HCD-MS2 and CID-MS3 modes after endoproteinase Glu-C digestion and boronate affinity chromatography. The abundance of glycation was assessed by protein labeling with (13)C(6)-glucose incubation. The analysis of native glycated CSF identified 111 glycation sites corresponding to 48 glycated proteins. Additionally, the in vitro high glucose level approach detected 265 glycation sites and 101 glycated proteins. The comparison of glycation levels under native and 15 mM glucose conditions showed relative concentration increases up to ten folds for some glycated proteins. This report revealed for the first time a number of key glycated CSF proteins known to be involved in neuroinflammation and neurodegenerative disorders. Altogether, the present study contains valuable and unique information, which should further help to clarify the pathological role of glycation in central nervous system pathologies. This article is part of a Special Issue entitled: Translational Proteomics.
Biomedical Proteomics Research Group, Department of Human Protein Sciences, University Medical Centre, University of Geneva, 1211 Geneva 4, Switzerland.
María
M
Ramírez-Boo
Feliciano
F
Priego-Capote
Alexandre
A
Hainard
Florent
F
Gluck
Pierre
P
Burkhard
Jean-Charles
JC
Sanchez
D000369:Aged, 80 and over; D002493:Central Nervous System Diseases / Q000134:cerebrospinal fluid*; D005947:Glucose / Q000378:metabolism; D006023:Glycoproteins / Q000134:cerebrospinal fluid*; D006031:Glycosylation; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*
D006023:Glycoproteins; D020543:Proteome; D005947:Glucose