Kolorimetrische mikromethode zur bestimmung des gesamteiweisses in eiweissarmen flüssigkeiten

Kaltwasser, F; Wolters, P; Pieper, Josef

doi:10.1016/0009-8981(67)90076-9

Cited by 45 publications

(15 citation statements)

References 6 publications

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“…For isolation of the intracellular glycosaminoglycans, the cell pellet was suspended in 0.5 ml per flask of 0.075M NaCl, and after 5 steps of freezing and thawing, a sample of the homogeneous suspension was removed for protein determination' 14 ', the remainder being processed as described above, including alkaline degradation, neutralization, centrifugation and dialysis.…”

Section: Methodsmentioning

confidence: 99%

Metabolism of Sulfated Glycosaminogtycans in Cultivated Bovine Arterial Cells. I. Characterization of Different Pools of Sulfated Glycosaminoglycans

Kresse¹,

Figura²,

Buddecke³

et al. 1975

Hoppe-Seyler´s Zeitschrift für physiologische Chemie

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Fibroblast-like" cells from the intimal pattern of chondroitin sulfate, dermatan sulfate, layer of bovine aorta were grown in culture. The heparan sulfate and hyaluronate. The distribution formation, composition, molecular weight and pattern of the extracellular glycosaminoglycans turnover rate of different pools of glycosaminoresembles closely that found in bovine aorta, glycans were investigated in cultures incubated in A small subfraction of the pericellular pool -tenthe presence of [ 35 S]sulfate or [ 14 C]glucosamine. tatively named "undercellular" pool -has been The newly synthesized glycosaminoglycans are characterized by its high heparan sulfate content, distributed into an extracellular pool (37 -58%), The intracellular and pericellular [ 35 S]glycosa cell-membrane associated or pericellular pool aminoglycan pools reach a constant radioactivity (23 -33%), and an intracellular pool (19 -30%), after 8 -12 h and 24 h, respectively, whereas the each pool exhibiting a characteristic distribution extracellular [ 35 S]glycosaminoglycans are seAbbreviations: CS, chondroitin sulfate; DS, dermatan sulfate; HA, hyaluronate; HS, heparan sulfate; ADi-4-S, 3-ö-A 4 -glucuronosyWV-acetylgalactosamine 4-sulfate; ADi-6-S, 3-O-A 4 -glucuronosyl-7V-acetylgalactosamine 6-sulfate; ADi-0-S, 3-0-A 4 -glucuronosyl-A r -acetylgalactosamine; PPO = 2,5-diphenyloxazol; POPOP = l,4-bis-4-methyl--5-phenyl-2-oxazolyl benzol. Enzymes: Chondroitin ABC lyase (EC 4.2.2.4): elimination of ADi-4-S, ADi-6-S, ADi-O-S from polysaccharides containing 1,4-0-hexosaminyl and l,3-j3-D-glucuronosyl or l,3,a-L-iduronosyl linkages, thus bringing about depolymerization. Acts on chondroitin 4-sulfate, chondroitin 6-sulfate and dermatan sulfate and slowly on hyaluronate. Chondroitin AC lyase (EC 4.2.2.5): elimination of ADi-4-S, ADi-6-S, ADi-O-S from polysaccharides containing l,4-(3-hexosaminyl and l,3-|3-D-glucuronosyl linkages, thus bringing about depolymerization. Acts on chondroitin 4-sulfate and chondroitin 6-sulfate, but less well on hyaluronate. Trypsin(EC3.4.21.4) NomenclatureIn this paper glycosaminoglycans are defined as [ 35 S]sulfate-or [ 14 C]hexosamine-containing polysaccharides characterized by the NaCl molarity by which they are eluted from Dowex 1X2 as compared with authentic glycosaminoglycan preparations and by their susceptibility towards chondroitinases. A covalent linkage of glycosaminoglycans to a protein or polypeptide moiety is not excluded. The term proteoglycan is used for gly cosaminoglycan-containing macromolecules which are excluded from Sepharose 4B gel or are eluted from it with a volume distinctly smaller than that of glycosaminoglycans.

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Section: Methodsmentioning

confidence: 99%

Metabolism of Sulfated Glycosaminogtycans in Cultivated Bovine Arterial Cells. I. Characterization of Different Pools of Sulfated Glycosaminoglycans

Kresse¹,

Figura²,

Buddecke³

et al. 1975

Hoppe-Seyler´s Zeitschrift für physiologische Chemie

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“…Equal activities from the resuspended precipitates and the supernatant of the controls were added to the culture medium of either mucopolysaccharidosis IIIB fibroblasts (deficient in a-N-acetylglucosaminidase) or Sandhoff fibroblasts (deficient in fl-Nacetylglucosaminidase). Enzyme endocytosis was determined as described by Ullrich et al (1978 (Kaltwasser et al, 1965) with bovine serum albumin as standard. Cell viability was monitored for each experimental condition by the Nigrosine test (Kaltenbach et al, 1958).…”

Section: Cell Culturementioning

confidence: 99%

An alternative hypothesis of cellular transport of lysosomal enzymes in fibroblasts. Effect of inhibitors of lysosomal enzyme endocytosis on intra- and extra-cellular lysosomal enzyme activities

Figura

Weber

1978

Biochemical Journal

109

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Recapture of lysosomal enzymes secreted by fibroblasts was inhibited by growing the cells in the presence of either free or immobilized antibodies against lysosomal enzymes or in the presence of phosphorylated carbohydrates known to interact with the cell-surface receptors for lysosomal enzymes. The following results were obtained. 1. Conditions that prevent recapture of released lysosomal enzymes increase the rate of extracellular accumulation of these enzymes up to twice that of controls. 2. Growing cells for 12 days in the presence of 0.5 mM-mannose 6-phosphate, which decreases ,B-N-acetylglucosaminidase endocytosis to less than 10% of that of controls, has no effect on the intracellular activity of this and four other lysosomal enzymes. 3. Growing cells for 4 days in the presence of 50mM-mannose 6-phosphate, which is a 1000-fold higher concentration than that required for 50% inhibition of lysosomal enzyme endocytosis, leads to a 4-fold increase in extracellular fl-N-acetylglucosaminidase accumulation and a decrease in intracellular enzyme. These results give evidence that, in fibroblasts, transfer of lysosomal enzymes into lysosomes does not require secretion before a receptor-mediated recapture [Hickman & Neufeld (1972) Biochem. Biophys. Res. Commun. 49,[992][993][994][995][996][997][998][999]. We propose that (a) lysosomal enzymes are present in a receptor-bound form in those vesicles that fuse with the cell membrane, (b) the major part of the lysosomal enzyme cycles via the cell surface in a receptor-bound form and (c) only a minor part of the lysosomal enzyme is released into the extracellular space during its life cycle.

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“…The ability to liberate N-acetylglucosamine 6-sulphate was drastically diminished in fibroblast homogenates from a patient with ,B-N-acetylhexosaminidase deficiency (Sandhoff disease) and lowered in ,6-galactosidase-deficient cells (GM, gangliosidosis). These data suggest that inOther methods Activity measurement of N-acetylgalactosamine 6-sulphate sulphatase and analyses of N-acetylglucosamine (Reissig et al, 1955), of other glucosaminoglycan constituents (Ludolph et al, 1981) and of cell protein (Kaltwasser et al, 1967) were performed as quoted earlier.…”

mentioning

confidence: 66%

Impaired degradation of keratan sulphate by Morquio A fibroblasts

Glössl¹,

Kresse²

1982

Biochemical Journal

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Upon incubation of keratan [35S]sulphate with normal fibroblasts both [35S]sulphate and N-acetylglucosamine 6-[35S]sulphate are liberated. From the products obtained after digestion with various mutant fibroblasts and with purified N-acetylgalactosamine 6-sulphate sulphatase we suggest that (i) [35S]sulphate is released almost exclusively from galactose 6-sulphate residues; (ii) N-acetylgalactosamine 6-sulphate sulphatase exhibits galactose 6-sulphate sulphatase activity; (iii) both sulphatase activities are deficient in Morquio disease type A.

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Kolorimetrische mikromethode zur bestimmung des gesamteiweisses in eiweissarmen flüssigkeiten

Cited by 45 publications

References 6 publications

Metabolism of Sulfated Glycosaminogtycans in Cultivated Bovine Arterial Cells. I. Characterization of Different Pools of Sulfated Glycosaminoglycans

Metabolism of Sulfated Glycosaminogtycans in Cultivated Bovine Arterial Cells. I. Characterization of Different Pools of Sulfated Glycosaminoglycans

An alternative hypothesis of cellular transport of lysosomal enzymes in fibroblasts. Effect of inhibitors of lysosomal enzyme endocytosis on intra- and extra-cellular lysosomal enzyme activities

Impaired degradation of keratan sulphate by Morquio A fibroblasts

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