title: Aberrant Glycosylation of Lumican in Aortic Valve Stenosis Revealed by Proteomic Analysis.
abstract: To identify proteins related to the pathophysiology of aortic valve stenosis (AS), we investigated the protein profiles of AS aortic valves. Specifically, proteins were extracted from a thickened and calcified area (AS-C) and an apparently non-thickened and non-calcified area (AS-N) in an identical aortic valve leaflet in each of 6 AS patients. The proteins were then separated by 2-dimensional gel electrophoresis (2DE). Protein spots detected by 2DE were compared between the AS-C and AS-N samples. Protein spots of interest were subjected to protein identification by mass spectrometry.In total, 670 protein spots were detected by 2DE, 28 of which showed more than 1.5-fold different intensity (P < 0.05) between the AS-C and AS-N samples. Proteins were identified in 17 out of the 28 spots. Fibrinogen and lumican were identified in 9 and 3 spots, respectively. Intensity of these 12 spots was lower in the AS-C samples than in the AS-N samples. In the 1D-Western blot analysis, 4 lumican bands (80 kDa, 75 kDa, 65 kDa, and 53 kDa) were detected, of which 2 bands with 80 kDa and 75 kDa showed lower intensity in the AS-C samples than in the AS-N samples. When de-glycosylated protein samples were used in the 1D-Western blot, only a single lumican band with ~40 kDa was detected, indicating that lumican was variously glycosylated and that highly glycosylated lumican molecules were decreased in AS-C.Collectively, insufficient glycosylation of lumican in the thickened and calcified areas of AS aortic valves may be involved in the pathophysiology of AS.
authors.affiliation: Department of Cardiovascular Surgery, St. Marianna University Graduate School of Medicine.
authors.forename: Hirotoshi
authors.initials: H
authors.lastname: Suzuki
authors.forename: Masahide
authors.initials: M
authors.lastname: Chikada
authors.forename: Michiyo K
authors.initials: MK
authors.lastname: Yokoyama
authors.forename: Manae S
authors.initials: MS
authors.lastname: Kurokawa
authors.forename: Takashi
authors.initials: T
authors.lastname: Ando
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Furukawa
authors.forename: Mitsumi
authors.initials: M
authors.lastname: Arito
authors.forename: Takeshi
authors.initials: T
authors.lastname: Miyairi
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Kato
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001024:Aortic Valve Stenosis / Q000378:metabolism*; D015153:Blotting, Western; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006031:Glycosylation; D006801:Humans; D007632:Keratan Sulfate / Q000378:metabolism*; D000071234:Lumican; D008297:Male; D008875:Middle Aged; D040901:Proteomics / Q000379:methods*
chemical_list: D011508:Chondroitin Sulfate Proteoglycans; C000604306:LUM protein, human; D000071234:Lumican; D007632:Keratan Sulfate