Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.2.1.36 (hyaluronidase)
 4,606 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The proteoglycans of cartilage are complex molecules in which chondroitin sulphate and keratan sulphate chains are covalently linked to a protein core, forming a polydisperse population of proteoglycan monomers. By interaction with hyaluronic acid and link proteins, the monomers form large macromolecular complexes. In vivo the proteoglycans mainly occur in such aggregates. In the electron microsope, the cartilaginous matrix can be seen to be made up of thin collagen fibrils and polygonal granules about 10-50 nm in diameter Addition of the polyvalent cationic dye Ruthenium Red to glutaraldehyde and osmium tetroxide fixatives yields a dense selective staining of the matrix granules. Following a short digestion of cartilage slices with either of the chondroitin sulphate-degrading enzymes hyaluronidase and chondroitinase or with the proteolytic enzyme papain, the matrix granules were few in number or completely absent and the proteoglycan content, measured as hexosamine, decreased by up to 90%. Similarly, extraction of the cartilage with 4 M guanidine-HCl removed all matrix granules and most of the proteoglycans. From these findings, it can be concluded that the matrix granules represent proteoglycans, most probably in aggregate form, and that Ruthenium Red staining may be used to study the distribution of these macromolecules in thin sections. As a complement to chemical studies on proteoglycan structure, it is also possible to observe and measure individual molecules in the electron microscope after spreading them into a monomolecular layer with cytochrome c. This technique has been applied in investigations on proteoglycans isolated from bovine nasal cartilage and other hyaline cartilages. The molecules in the monomer fractions appeared as an extended central core filament to which about 25--30 side-chain filaments were attached at various intervals. The core filament, averaging about 300 nm in length, was interpreted as representing the polysaccharide binding part of the protein core and the side-chain filaments, averaging about 45 nm in length, as representing the clusters of chondroitin sulphate chains. Statistical treatment of the collected data indicated that no distinct subpopulations existed within the monomer fractions. The electron microscopic results correlated well with chemical data for the corresponding fractions and together with recent observations on various aggregate fractions strongly support present concepts of proteoglycan structure.
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PMID:Electron microscopy of cartilage proteoglycans. 6 24

This paper reports an unrecognized aspect of phosphotungstic acid staining at low pH. It provides an on-section staining method in which sialic acid-containing molecules can be demonstrated in the laminae rarae of the rat glomerular basement membrane. The staining in the basement membrane became negative after perfusion with the following cations: protamine sulphate, hexadimethrine, Alcian Blue, Ruthenium Red and Toluidine Blue. Blocking was not achieved with Alcian Blue at about pH 1. The staining was also abolished after mild methylation and demethylation restored the contrast. This is suggestive of the involvement of carboxyl groups. Prior digestion with pronase, trypsin and neuraminidase rendered the laminae rarae negative, whereas hyaluronidase, chondroitinase ABC and crude heparinase were without effect. This indicates that sialic acid groups are detected by this method and that heparan sulphate does not interfere. The staining of the epithelial plasma membrane, also carrying sialic acid groups, remained positive after neuraminidase treatment. It is presumed that this method can be applied successfully for detecting changes in the sialic acid content of the laminae rarae in rat glomerular basement membranes under normal and pathological conditions.
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PMID:Demonstration of sialic acid groups in the glomerular basement membrane of the rat with phosphotungstic acid at low pH. 241 Mar 95

The presence of hyaluronate in the capsular space of the cat muscle spindle was demonstrated using alcian blue staining at various pHs, the critical electrolyte concentration technique and hyaluronidase treatment. In spindles with intact capsules an extracellular marker, the dye Ruthenium Red, gained access to the capsular space through the gap in the sleeve region, but for a limited distance. In muscle spindles with the capsule nicked, the marker diffused into the capsular space in the equatorial region, revealing a dense network in this space which consisted of globular structures interconnected by thin filaments. Based on their thickness, these filaments were inferred to be hyaluronic acid, and the globular structures were inferred to be protein molecules. Longitudinal diffusion of the dye into the capsular space through the nicked site was limited. The limited diffusion is probably due to electrostatic binding of the dye, which is a hexavalent cation, to negatively charged glycosaminoglycan hyaluronate that is present in the space. The transcapsular potential was measured by use of glass micropipettes filled with 3 M-KCl. The value was 15 mV +/- 4 (average +/- S.D., n = 12; range, 10-20 mV) inside negative. The input resistance and capacitance of the capsule, measured with two independent electrodes, varied widely (1.3-8.0 M omega and 0.5-1.3 nF, n = 4) and the capsule showed marked delayed rectification to outward current pulses. [K+] in the space measured with K+-sensitive resin-filled glass micropipettes was a few millimolar higher than that in the bathing solution. The effects of [K+] and [Ca2+] on impulse activities were examined in spindles with intact capsules or with partially resected capsules. In spindles with intact capsules the effects of [K+] and [Ca2+] were significantly less or negligible compared with those in spindles with the capsule opened. Hyaluronidase (approximately 10(-4) g/ml) added to the bathing solution around nicked capsules significantly reduced both resting and stretch-induced impulse activities in 40-50 min. By this time the capsular space was completely collapsed. An increase in [K+] of the bathing solution from 3.5 to 6 or 8 mM restored these impulse activities. A similar restoring effect was also observed when [Ca2+] in the bathing solution was reduced.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Studies of capsule and capsular space of cat muscle spindles. 294 10

Ultrastructural studies on human lung were performed with special attention to the interstitial acid mucopolysaccharides by Ruthenium Red staining and several enzyme digestion tests with Streptomyces hyaluronidase, chondroitinase ABC, chondroitinase AC, heparinase, trypsin and collagenase. Periodic lateral granules on the major cross bands of collagen fibrils and amorphous coats on them became visible by Ruthenium Red staining. The surface of elastic fibres, associated microfibrils, and some fine fibrils 10-20 nm in diameter were stained. Ruthenium Red also stained the surface of fibroblast and smooth muscle cells, basement membrane and filamentous long segments. In the interstructural space, granular substances 10-80 nm in diameter and fine filaments 3--4 nm thick, which formed a fine reticular network, were clearly observed. They were not visible on the usual thin section. The granular substances were located on the cross points of the fine filaments. They spread continuously and connected with each of the cells and extracellular structures in the pulmonary interstitium. The results of the enzyme digestion tests on the Ruthenium Red-positive material are discussed.
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PMID:Electron microscopic observations on pulmonary connective tissue stained by Ruthenium Red. 617 14