Modifications of a specific assay for hydroxyproline in urine

Kivirikko, Kari I.; Laitinen, Ossi; Prockop, Darwin J.

doi:10.1016/0003-2697(67)90160-1

Cited by 1,035 publications

(317 citation statements)

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“…This problem can be overcome by using a large quantity of a synthetic peptide substrate and by measuring the amount of hydroxyproline synthesized by a chemical procedure [34]. To study whether the changes in prolyl hydroxylase activity found here were real or due to an artefact, the enzyme activity was assayed in the same early-log and late-log cells by two different procedures, and identical increases were found (Table 1).…”

Section: Changes In Enzyme Activities In Cultured 3t6 Cellsmentioning

confidence: 94%

Intracellular Enzymes of Collagen Biosynthesis in 3T6 Fibroblasts and Chick‐Embryo Tendon and Cartilage Cells

Risteli

Salo

et al. 1979

European Journal of Biochemistry

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An increase of about 4-9-fold was found in prolyl and lysyl hydroxylase activities per cell when 3T6 fibroblasts were grown from the early-log to the stationary phase, whereas essentially no changes were found in hydroxylysyl galactosyltransferase and galactosylhydroxylysyl glucosyltransferase activities. A control experiment using a different assay procedure indicated that the increase in prolyl hydroxylase activity was not due to any methodological artefact. The results demonstrate for the first time that these four intracellular enzymes of collagen biosynthesis are not regulated in an identical manner within one cell type.Freshly isolated chick embryo cartilage cells, which synthesize type I1 collagen, had higher lysyl hydroxylase and hydroxylysyl galactosyltransferase activities than freshly isolated chick embryo tendon cells, whereas no difference was found between them in prolyl hydroxylase and galactosylhydroxylysyl glucosyltransferase activities. The greater extent of the hydroxylation of lysyl residues and galactosylation of hydroxylysyl residues in type I1 than in type I collagen is thus probably due in part to a higher activity of the corresponding enzymes in the cartilage cells, but the greater extent of the glucosylation of galactosylhydroxylysyl residues cannot be explained by any higher activity of the glucosyltransferase.Prolyl and lysyl hydroxylase activities were markedly lower in the cultured 3T6 cells than in freshly isolated chick embryo tendon cells, particularly when expressed per unit solubilized cell protein. These data agree with the lower rate of collagen production by the 3T6 cells. Unexpectedly, the two hydroxylysyl glycosyltransferase activities were somewhat higher in the 3T6 cells than in the tendon cells when expressed per cell, a finding which may explain the pronounced glycosylation of the collagen synthesized by the 3T6 cells.On incubation of the 3T6 cells with 5 pg/ml puromycin for 10 h the intracellular enzyme activities of collagen biosynthesis decreased by about 15 -30 in the early-log phase and by about 10-20 x> in the late-log phase. The results suggest that the half-lives of all four enzyme activities are at least about 24 h in the early-log phase and possibly even longer in the late-log phase. ~~~ ~ ~~~f37:i.mes and h'ommclrrure. Prolyl 4-hydroxylase or prolylglycyl-peptide, 2-oxoglutarate: oxygen oxidoreductase (4-hydroxylating) (EC 1.14.1 1.2) has usually been termed prolyl hydroxylase; recently, however, two separate enzymes, prolyl 4-hydroxylase and prolyl 3-hydroxylase have been demonstrated [4]; the present work concerns only the 4-hydroxylase activity, and the term prolyl hqdroxylasc is used exclusively to denote this.The term lysyl hydroxylase is used for peptidyllysine, 2-oxog1utarate:oxygen 5-oxidoreductase (EC 1.14.11.4). The two hydroxylysyl glycosyltransferases. UDPgalactose : 5-hydroxylysine-collagen galactosyl transferase (EC 2.4.1.50) and UDPglucose: 5-hydroxylysine-collagen glucosyltransferase (EC 2.4.1.66) have earlier been known as collagen galactosy...

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Section: Changes In Enzyme Activities In Cultured 3t6 Cellsmentioning

confidence: 94%

Intracellular Enzymes of Collagen Biosynthesis in 3T6 Fibroblasts and Chick‐Embryo Tendon and Cartilage Cells

Risteli

Salo

et al. 1979

European Journal of Biochemistry

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“…Total collagen was determined by hydrolyzing liver samples in 6 N HCl and by measuring hydroxyproline according to the method of Kivirikko et al 26 Collagen content was estimated by multiplying the amount of hydroxyproline by a factor of 7.69. Results are expressed as milligram of collagen per gram of liver (wet weight).…”

Section: Hydroxyproline Contentmentioning

confidence: 99%

F2-isoprostanes stimulate collagen synthesis in activated hepatic stellate cells: a link with liver fibrosis?

Comporti

Arezzini

Signorini

et al. 2005

Laboratory Investigation

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Carbon tetrachloride (CCl 4 )-induced hepatic fibrosis has been considered to be linked to oxidative stress and mediated by aldehydic lipid peroxidation products. In the present study, we investigated whether collagen synthesis is induced by F 2 -isoprostanes, the most proximal products of lipid peroxidation and known mediators of important biological effects. By contrast with aldehydes, F 2 -isoprostanes act through receptors able to elicit definite signal transduction pathways. In a rat model of CCl 4 -induced hepatic fibrosis, plasma F 2 -isoprostanes were markedly elevated for the entire experimental period; hepatic collagen content also increased. When hepatic stellate cells (HSCs) from normal liver were cultured with F 2 -isoprostanes in the concentration range found in the in vivo studies (10 À9 -10 À8 M), a striking increase in DNA synthesis (reversed by the thromboxane A 2 antagonist SQ 29 548), in cell proliferation and in collagen synthesis was observed. Total collagen content was similarly increased. Moreover, F 2 -isoprostanes markedly increased the production of transforming growth factor-b1 by U937 cells, considered a model of liver macrophages. The data provide evidence for the possibility that F 2 -isoprostanes generated by lipid peroxidation in hepatocytes mediate HSC proliferation and collagen production seen in hepatic fibrosis.

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“…The hydroxyproline content was measured, as described elsewhere. 32 The liver-specific cytosolic enzyme activities of glutamic-oxaloacetic transaminase, glutamic-pyruvate transaminase, and alkaline phosphatase in sera were analyzed using an automated-analyzer (Hitachi Co., Tokyo, Japan). Plasma fibrinogen concentrations and hepaplastin tests were examined using commer- HGF.…”

mentioning

confidence: 99%

Preventive and therapeutic effects in rats of hepatocyte growth factor infusion on liver fibrosis/cirrhosis

et al. 1997

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Liver cirrhosis is characterized by the hyper-accumulation Liver fibrosis/cirrhosis is characterized by hyper-accumulaof connective tissue components in the liver and is often tion of fibrous tissue components and is commonly observed accompanied by hypo-albuminemia, ascites, and esophageal in later or terminal states of chronic hepatic diseases. In ongovarices. In a prolonged stage, severe hepatic failure, or hepaing work, we found that the administration of human recombitocellular carcinoma, can occur. Chronic hepatic injuries nant hepatocyte growth factor (hrHGF) suppressed the onset caused by drugs, chemicals, alcohol abuse, or viral infections of liver fibrosis/cirrhosis in several distinct models and accelare predominant pathogenic causes for cirrhosis. Although erated the recovery from liver fibrosis/cirrhosis in rats. Rethe pathogenic mechanisms of cirrhosis are not fully underpeated administration of porcine serum for 10 weeks to rats stood, the over-production of extracellular matrices in the induced liver fibrosis without any accompanying hepatocelluliver, as well as of chronic parenchymal hepatocellular injury, lar injuries; in addition, the intravenous (i.v.) administration are likely to initiate the onset of cirrhosis. Therefore, preferof hepatocyte growth factor (HGF) to these rats suppressed ential hepatocellular replication and cytoprotection against increases in fibrous components and hydroxyproline contents hepatic injuries, as well as the stimulation of degradation in the liver, thus preventing the onset of liver fibrosis. Reand remodeling of extracellular fibrous components, seems peated administration of dimethylnitrosamine (DMN) for four to inhibit the onset or progress of hepatic fibrosis/cirrhosis. weeks induced liver cirrhosis, as characterized by the hyperSeveral lines of studies have shown that hepatocyte growth accumulation of fibrous components, infiltration of mononufactor (HGF), originally purified as a potent mitogen for rat clear leukocytes, and hepatic dysfunction. When HGF was hepatocytes, 1-3 is the long-sought hepatotrophic factor for injected daily for four weeks along with DMN-treatment, the liver regeneration. [4][5][6][7] Following the onset of various types of onset of DMN-induced hepatic fibrosis/cirrhosis was suphepatic injuries, HGF messenger RNA expression is rapidly pressed; the numbers of infiltrating mononuclear cells, fibrous tissue components, and hydroxyproline content in the liver up-regulated in the livers of experimental animals. 8,9 Serum were decreased. When HGF was injected for two weeks fol-HGF levels are elevated in patients with various hepatic dislowing four weeks of DMN-treatment, HGF accelerated the orders. 10,11 Extensive in vivo studies on the efficacy of recomrecovery from liver cirrhosis and prevented death due to he-binant HGF on hepatic regeneration revealed that HGF elicits patic dysfunction. Likewise, HGF-injection suppressed the on-a potent hepatotrophic action. HGF strongly stimulates DNA set of liver fibrosis, when liver fibrosis had be...

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Modifications of a specific assay for hydroxyproline in urine

Cited by 1,035 publications

References 7 publications

Intracellular Enzymes of Collagen Biosynthesis in 3T6 Fibroblasts and Chick‐Embryo Tendon and Cartilage Cells

Intracellular Enzymes of Collagen Biosynthesis in 3T6 Fibroblasts and Chick‐Embryo Tendon and Cartilage Cells

F2-isoprostanes stimulate collagen synthesis in activated hepatic stellate cells: a link with liver fibrosis?

Preventive and therapeutic effects in rats of hepatocyte growth factor infusion on liver fibrosis/cirrhosis

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