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Y Oike

Publications and source records attributed to Y Oike.

At least 73 records · Page 4Linked to original sources
PubMed1984

Characteristics of the core protein of the aggregating proteoglycan from the Swarm rat chondrosarcoma.

A ternary complex of hyaluronic acid-binding region and link protein bound to hyaluronic acid was isolated from limit clostripain digests of proteoglycan aggregates isolated from the Swarm rat chondrosarcoma. Under these conditions, the hyaluronic acid-binding region has a molecular weight of approximately equal to 65,000 (HA-BR65). N-terminal amino acids in the complex were selectively 14C-carbamylated. The resulting derivatized HA-BR65 was isolated, and tryptic peptide maps were prepared and developed on two-dimensional TLC sheets. A single, labeled peptide was obtained which gave a Mr by approximately equal to 8,000 by SDS-PAGE. Chymotrypsin digestion of the ternary complex reduced the molecular weight of HA-BR65 to a polypeptide of approximately equal to 55,000 (HA-BR55) which still retains the same N-terminal tryptic peptide. Partial digestion of proteoglycan aggregates with clostripain generated a series of larger intermediates with the hyaluronic acid-binding region. Direct SDS-PAGE analysis revealed one major intermediate with approximately equal to 109,000 (HA-BR109) as well as HA-BR65. After chondroitinase digestion, two additional prominent intermediates were observed on a SDS-PAGE gel at Mr approximately equal to 120,000 (HA-BR120) and approximately equal to 140,000 (HA-BR140). All the intermediates were recognized by a monoclonal antibody specific for the hyaluronic acid-binding region, and all of them contained the same N-terminal tryptic peptide. The results indicate that the N terminus of the core protein is at the hyaluronic acid-binding end of the proteoglycan and that the chondroitin sulfate chains are first present on the core protein in a region between 109,000 and 120,000 molecular weight away from the N terminus.

Amino Acid Sequence↗
PubMed1984

Appearance of distinct types of proteoglycan in a well-defined temporal and spatial pattern during early cartilage formation in the chick limb.

Our recent studies have shown that chick embryo epiphyseal cartilage synthesizes three distinct species of proteoglycan (PG-H, PG-Lb, and PG-Lt) which are analogous in having glycosaminoglycan side chains of the chondroitin (dermatan) sulfate type but different from one another in regard to the structure of core protein. In the present report, the expression of PG-H and PG-Lb has been studied in developing chick hind limbs (stages 19-33), using antibodies specific for these substances in indirect immunofluorescence. At the onset of cartilage morphogenesis (stage 24), PG-H became recognizable in the cartilage primordia, whereas a parallel section stained for PG-Lb showed no reaction. The first evidence of PG-Lb appearance was seen in a stage 28 cartilage (e.g., tibia) in which the cells in the middiaphysis became elongated in a direction perpendicular to the long axis of the cartilage. The PG-Lb fluorescence was confined to the zone of these flattened, disc-like cells, whereas the fluorescence for PG-H was uniformly distributed throughout the cartilage. With further development of cartilage (stage 29 approximately), the zone of flattened cells spread proximally and distally, and simultaneously large hypertrophied cells appeared at the diaphyseal region. During these zonal changes of cell morphology, the PG-Lb fluorescence remained restricted to the zone of flattened cells. Parallel sections stained for PG-H, in contrast, showed an evenly distributed pattern of the PG-H fluorescence throughout the cartilage. The results indicate that the appearance of PG-Lb is closely associated with the zonal changes of cell shape and orientation along the proximal-distal axis of the developing limb cartilage, and further suggest that the flattened chondrocytes in this particular zone have undergone additional changes in gene expression to form an extracellular matrix of still another chemical property.

Animals↗
PubMed1983

The occurrence of three different proteoglycan species in chick embryo cartilage. Isolation and characterization of a second proteoglycan (PG-Lb) and its precursor form.

Three different molecular species of proteoglycan (designated PG-H, PG-Lb, and PG-Lt) have been isolated from chick embryo epiphyseal cartilage. PG-H is a major proteoglycan of the tissue and identical, or nearly identical, with so-called cartilage-characteristic proteoglycan previously described in mammalian and avian cartilages. The third proteoglycan, PG-Lt, differs from the other two in containing disulfide-bonded collagenous polypeptides (Noro, A., Kimata, K., Oike, Y., Shinomura, T., Maeda, N., Yano, S., Takahashi, N., and Suzuki, S. (1983) J. Biol. Chem. 258, 9323-9331). The second proteoglycan, PG-Lb, consists of a core protein with Mr congruent to 52,000 dermatan sulfate copolymer chains with glucuronic acid/iduronic acid residues. Upon chondroitinase ABC digestion, the proteoglycan yields a protein-enriched core fraction of Mr congruent to 43,000. Its amino acid composition, tryptic peptide profile, and immunochemical properties indicate that PG-Lb is distinctly different from PG-H and PG-Lt in core protein structure. PG-Lb shows no specific binding with hyaluronic acid. Pulse-chase experiments with [3H]serine indicate that PG-Lb is first synthesized as a precursor form (pro-PG-Lb) that can be distinguished from PG-Lb by the production of a core molecule of Mr congruent to 52,000 after chondroitinase ABC digestion. This core molecule is labeled when [2-3H]mannose is used as a precursor, suggesting that it contains a glycoprotein type oligosaccharide. Since the core molecule from pro-PG-Lb is significantly larger in molecular weight than that from PG-Lb, the conversion of pro-PG-Lb to PG-Lb should involve scission of the polypeptide or possibly removal of mannose-containing oligosaccharide chain.

Amino Acids↗
PubMed1983

Isolation and characterization of a third proteoglycan (PG-Lt) from chick embryo cartilage which contains disulfide-bonded collagenous polypeptide.

Chick embryo epiphyseal cartilage has been shown to contain three different proteoglycan species (PG-H, PG-Lb, and PG-Lt). This report is concerned with the purification and characterization of the third proteoglycan, PG-Lt. The proteoglycan can be separated from the other two by virtue of its low buoyant density in a CsCl density gradient and further purified by consecutive ion exchange and gel chromatography. The final preparation is composed of PG-Lt monomer and PG-Lt oligomer. The amino acid composition of PG-Lt is quite different from that of PG-H and PG-Lb and rather resembles that of collagens with respect to high content of glycine and high degrees of hydroxylation of proline and lysine. PG-Lt monomer is composed of disulfide-bonded subunits of Mr congruent to 120,000 and 190,000 as demonstrated by its gel electrophoretic behavior after reduction with 2-mercaptoethanol. The latter, but not the former, contains dermatan sulfate chains with glucuronic acid/iduronic acid residues and yields a protein-enriched core molecule of Mr congruent to 100,000 after digestion with chondroitinase ABC. Both of the protein subunits are completely digestible with bacterial collagenase. Immunofluorescence microscopic examination of cartilage tissues, using an antibody against PG-Lt, shows that this proteoglycan exists in both the cartilage matrix and perichondrial noncartilagenous region. When chondrocytes are plated onto tissue culture dishes, the antibody stains strands found on the cell surfaces and in the intercellular space of substrate-attached cell layers, suggesting that PG-Lt mediates cell-to-cell and cell-to-substrate contacts.

Amino Acids↗
PubMed1983

[Clinical estimation of the relation between coronary artery disease and regional myocardial blood flow by continuous thermodilution with the multithermistor catheter].

To evaluate coronary hemodynamics more precisely in ischemic heart disease, great cardiac vein flow ( GCVF ) and coronary sinus flow (CSF) were simultaneously measured before and during atrial pacing by a continuous thermodilution technique with the multithermistor catheter in 10 patients with significant coronary stenosis (narrowing of the left anterior descending artery of 75% or more) (Group 1: CAD) and 12 cases with the normal coronary artery (narrowing of 25% or less) (Group 2: NonCAD ). GCVF reflects the anterior regional flow of the left ventricle and CSF reflects total left ventricular flow. Between the two groups, no significant differences were noticed in either resting GCVF or resting CSF. However, atrial pacing induced a reduction of coronary resistance (CR) and increasing regional myocardial blood flow in both groups in the same way, and the maximal pacing provoked anginal episode in CAD group in which both the percent decrease of coronary resistance (CR) (% delta CRant 25.3% and % delta CRt 23.3%) and the percent increase of regional myocardial blood flow (% delta GCVF 49.6% and % delta CSF 40.6%) were lower than those in NonCAD group (% delta CRant 42.3%, % delta CRt 37.6%, % delta GCVF 80.6%, and % delta CSF 68.3%, respectively). These data imply that resting coronary blood flow may be maintained as normal in patients with angiographically significant coronary stenosis, though coronary reserve probably decreased in these patients. In this respect, the continuous thermodilution technique using the multithermistor catheter may be useful for the clinical estimation of coronary hemodynamics induced by various interventions. This technique can be performed repeatedly for a short time and permits simultaneous assessment of venous blood flow draining from the anterior wall and of total left ventricular flow.

Cardiac Pacing, Artificial↗
PubMed1982

A mapping technique for probing the structure of proteoglycan core molecules.

Our previous work showed that treatment of chick embryo cartilage proteoglycan (PG-H) with chondroitinase-AC II and keratanase yielded a protein-rich core fraction having enzymatically modified linkage oligosaccharides. The core sample has now been analyzed by tryptic peptide mapping, in which the isolated core sample contained in a single Coomassie blue-staining band from a dried slab gel is radioiodinated and treated with trypsin, and the resultant tryptic peptides are displayed two-dimensionally on a silica gel thin layer plate. The map thus obtained exhibited 22 major peptide spots, the resolution and location of which were reproducible. In order to identify regions of the core polypeptide from which the tryptic peptides are derived, PG-H was cleaved with clostripain under conditions that yield a hyaluronic acid-binding fragment with an apparent Mr = 150,000 and chondroitin sulfate-peptide clusters of smaller molecular sizes. Although the peptide maps of the two size classes of clostripain fragments differed significantly from each other, the patterns of spots, as a whole, were extensively similar to those observed with the intact core molecule. These results have provided additional evidence that PG-H has a single, nonvariable core protein structure. In addition, the technique used here will provide a versatile method for the identification of genetic types in this increasingly complex family of matrix macromolecules.

Animals↗
PubMed1980

Structural analysis of chick-embryo cartilage proteoglycan by selective degradation with chondroitin lyases (chondroitinases) and endo-beta-D-galactosidase (keratanase).

Digestion of chick-embryo cartilage proteoglycan (type H) with chondroitin AC II lyase or keratanase, in the presence of EDTA, N-ethylmaleimide, phenylmethanesulphonyl fluoride and pepstatin, resulted in the removal of the bulk of the chondroitin sulphate or keratan sulphate chains respectively, without altering the protein portion of the macromolecule. An exhaustive treatment of the proteoglycan with chondroitin AC II lyase followed by digestion with keratanase yielded a core fraction having the enzymically modified linkage oligosaccharides. Zonal sedimentation of this core preparation on a sucrose gradient in 0.5% SDS resulted in a single narrow band with a sedimentation coefficient of 6S. In 4 M-guanidinium chloride, the core preparation showed a tendency to aggregate to multiple-molecular-weight forms which could dissociate in the presence of Triton X-100. The results indicate that the preponderance of glycosaminoglycans in the proteoglycan molecule is a main reason for both polydispersity and hydrophilicity of the proteoglycan preparation, and further suggest that the enzymic procedures could prove useful as a method to obtain new information about the structure and properties of proteoglycan core molecules.

Animals↗
PubMed1980

Proteinase activity in chondroitin lyase (chondroitinase) and endo-beta-D-galactosidase (keratanase) preparations and a method to abolish their proteolytic effect on proteoglycan.

Significant amounts of proteinase activity have been found in chondroitin ABC lyase (EC 4.2.2.4), chondroitin AC II lyase and endo-beta-D-galactosidase (keratanase) from commercial sources. It would appear, therefore, that certain earlier biochemical and histochemical studies, which employed these commercial enzyme preparations for their presumed ability to degrade only glycosaminoglycans, may require re-evaluation. A mixture of EDTA, N-ethylmaleimide, phenylmethanesulphonyl fluoride and pepstatin abolishes the effect of the contaminating proteinases on proteoglycan with less significant effect on the chondroitin lyase or keratanase activity.

Animals↗
PubMed1979

A clinical study on urinary kallikrein in patients with renal diseases.

The urinary kallikrein activity (KA) was measured to investigate its significance in the renal diseases by using tosyl-arginine methyl ester (TAME) as a substrate. The examinees were 94 patients with renal diseases and 25 normal persons. The daily urinary kallikrein excretion (KE, KE=KAxdaily urinary volume) is less in chronic glomerulonephritis and outstandingly less in chronic renal failure than in the normal controls. The KE also shows a positive correlation moderately to 15-min PSP excretion and relatively to creatinine clearance. KE is closely related to renal function and decreases with the degree of renal damage. KA has no relation to the concentration of urine protein, but it was parallel, in general, to the urokinase activity. In nephrotic syndrome, KA tends to show a negative correlation to the urinary alpha 1-antitrypsin. alpha 1-antitrypsin may have a function as an inhibitor to the urinary kallikrein.

Adolescent↗
PubMed1968

[Meningeal tumor].

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Brain Neoplasms↗