Open AccessJournal Article
Immunoanalysis of keratan sulfate proteoglycan from corneal scars.
TLDR
Corneal keratan sulfate proteoglycan from scar tissue of experimental penetrating corneal wounds in rabbits was analyzed 2-8 weeks after injury using three previously characterized antibodies and showed identical quantitative binding of antibodies against core protein and KS from normal and scar tissue.Abstract:
Corneal keratan sulfate proteoglycan (KSPG) from scar tissue of experimental penetrating corneal wounds in rabbits was analyzed 2-8 weeks after injury using three previously characterized antibodies. Keratan sulfate (KS) was identified in 2 week scars and normal corneal tissue by indirect immunofluorescence using a monoclonal antibody against sulfated KS epitopes. KSPG was measured in unfractionated extracts of scar and of normal corneal tissue using a "sandwich" enzyme-linked immunosorbent assay (ELISA). In extracts of 2 week scars, KSPG molecules reacting with two different anti-KS monoclonal antibodies were 55% and 82% as abundant as in normal tissue extracts. Ion exchange high performance liquid chromatography (HPLC) of tissue extracts found qualitatively similar elution profiles of KSPG antigens from both scar and normal tissues. Direct ELISA of the HPLC-purified KSPG showed identical quantitative binding of antibodies against core protein and KS from normal and scar tissue. KS in the HPLC-purified extracts was sensitive to digestion with endo-beta-galactosidase, whereas core protein antigens were not affected by this enzyme, as expected. Alteration of the antigenic characteristics of the KSPG of scars was detected with a competitive immunoassay using immobilized monoclonal antibodies against KS. KS in extracts from 2, 6, and 8 week scars competed only 5-11% as effectively as KS from normal cornea, although core protein antigens in the scar extracts competed 61-80% as well as those of normal cornea.(ABSTRACT TRUNCATED AT 250 WORDS)read more
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Keratocyte Phenotype Mediates Proteoglycan Structure A ROLE FOR FIBROBLASTS IN CORNEAL FIBROSIS
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Roles of lumican and keratocan on corneal transparency.
Winston W.-Y. Kao,Chia-Yang Liu +1 more
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Novel aspects of the ultrastructural organization of human corneal keratocytes.
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Mimecan/osteoglycin-deficient mice have collagen fibril abnormalities.
Elena S. Tasheva,Anja Koester,Avelina Q. Paulsen,Alexander S. Garrett,Daniel L. Boyle,Harriet J. Davidson,Min Song,Niles Fox,Gary W. Conrad +8 more
TL;DR: It is demonstrated that mimecan, similar to other members of the SLRP gene family, has a role in regulating collagen fibrillogenesis in vivo and further studies, such as functional challenges, an evaluation of potential compensation by other proteins, and generation of double-knockouts will be necessary to fully uncover physiological functions of mime can in mice.
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The Use of X-ray Scattering Techniques to Determine Corneal Ultrastructure
Keith M. Meek,Andrew J. Quantock +1 more
TL;DR: The ideas that underpin the use of X-ray diffraction to investigate corneal ultrastructure are explained and how the knowledge gained to date has far-reaching implications for tissue biomechanics, disease changes and transparency are shown.
References
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A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
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Journal ArticleDOI
3,3',5,5' - Tetramethylbenzidine as an Ames test negative chromogen for horse-radish peroxidase in enzyme-immunoassay.
TL;DR: The use of 3,3',5,5' - tetramethylbenzidine as non-mutagenic chromogen for the end point determination in enzyme-immunoassay (EIA) is described, which led to an improvement of sensitivity and precision of the assays and makes EIA even more competitive with other types of immunoassays.
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Identification of a monoclonal antibody that specifically recognizes corneal and skeletal keratan sulfate. Monoclonal antibodies to cartilage proteoglycan
TL;DR: The 1/20/5-D-4 monoclonal antibody appears to recognize a common determinant in their polysaccharide moieties, consistent with several biochemical analyses showing the absence of keratan sulfate in proteoglycan synthesised by this tissue.
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Quantification of keratan sulfate in blood as a marker of cartilage catabolism.
Eugene J.-M.A. Thonar,Mary Ellen Lenz,Gordon K. Klintworth,Bruce Caterson,Lauren M. Pachman,Paul B. Glickman,Robert S. Katz,John P. Huff,Klaus E. Kuettner +8 more
TL;DR: If the appearance of elevated levels of serumKS do indeed correlate with the extent of cartilage erosion or destruction in individuals with OA, measurements of serum KS levels will prove extremely useful in the assessment and diagnosis of this joint disease.
Journal ArticleDOI
The core proteins of large and small interstitial proteoglycans from various connective tissues form distinct subgroups
Dick Heinegård,A Björne-Persson,L Cöster,A Franzén,S Gardell,Anders Malmström,Mats Paulsson,R Sandfalk,K G Vogel +8 more
TL;DR: Large and small proteoglycans were separately isolated from a number of connective tissues and compared to determine the extent of structural similarity, and two groups can be identified, although amino acid composition and protein core sizes are grossly similar.