کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10913877 | 1088643 | 2010 | 13 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Molecular interactions in the retinal basement membrane system: A proteomic approach
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کلمات کلیدی
mAbSDS PAGEHSPGGAGIPIILMemPAI - EMLC-MS/MS - LC-MS / MSMonoclonal antibody - آنتی بادی مونوکلونالsodium dodecylsulfate polyacrylamide gel electrophoresis - الکتروفورز ژل پلی اکریللید سدیم دودسیل سولفاتexponentially modified protein abundance index - شاخص فراوانی پروتئین پروتئین تغییر یافتهInternational Protein Index - شاخص پروتئین بین المللیMass spectrometry - طیف سنجی جرمیinner limiting membrane - غشای محدود کننده درونیbasement membrane - غشای پایهLaminin - لامینینYoung's modulus - مدول یانگHeparan sulfate proteoglycan - هپارین سولفات پروتئگلیکانProteomics - پروتئومیکسCollagen - کلاژنGlycosaminoglycan - گلیکوزآمینوگلیکان
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
تحقیقات سرطان
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چکیده انگلیسی
Basement membranes (BMs) are physiologically insoluble extracellular matrix sheets present in all multicellular organisms. They play an important role in providing mechanical strength to tissues and regulating cell behavior. Proteomic analysis of BM proteins is challenged by their high molecular weights and extensive post-translational modifications. Here, we describe the direct analysis of an in vivo BM system using a mass spectrometry (MS) based proteomics approach. Retinal BMs were isolated from embryonic chick eyes. The BM macromolecules were deglycosylated and separated by low percentage gradient SDS PAGE, in-gel digested and analyzed by LC-MS/MS. This identified over 27 extracellular matrix proteins in the retinal BM. A semi-quantitative measure of protein abundance distinguished, nidogens-1 and -2, laminin subunits α1, α5, β2, and γ1, agrin, collagen XVIII, perlecan, FRAS1 and FREM2 as the most abundant BM protein components. Laminin subunits α3, β1, γ2, γ3 and collagen IV subunits α5 and α6 were minor constituents. To examine binding interactions that contribute to the stability of the retinal BM, we applied the LC-MS/MS based approach to detect potential BM complexes from the vitreous. Affinity-captured nidogen- and heparin-binding proteins from the vitreous contained > 10 and > 200 proteins respectively. Comparison of these protein lists with the retinal BM proteome reveals that glycosaminoglycan and nidogen binding interactions play a central role in the internal structure and formation of the retinal BM. In addition, we studied the biomechanical qualities of the retinal BM before and after deglycosylation using atomic force microscopy. These results show that the glycosaminoglycan side chains of the proteoglycans play a dominant role in regulating the thickness and elasticity of the BMs by binding water to the extracellular matrix. To our knowledge, this is the first large-scale investigation of an in vivo BM system using MS-based proteomics.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Matrix Biology - Volume 29, Issue 6, July 2010, Pages 471-483
Journal: Matrix Biology - Volume 29, Issue 6, July 2010, Pages 471-483
نویسندگان
Manimalha Balasubramani, Emanuel M. Schreiber, Joseph Candiello, G.K. Balasubramani, Justin Kurtz, Willi Halfter,