Protocollagen Proline Hydroxylase Activity Research Articles

Synovial fluid beta 2 microglobulin and hydroxyproline fractions in rheumatoid arthritis and nonautoimmune arthropathies.

Fourteen patients with classical and definite seropositive rheumatoid arthritis (RA), 5 patients with microcrystalline arthritis, and 7 patients with osteoarthrosis were studied with respect to markers of newly synthesised collagen (synovial NDOH pro levels); markers of connective tissue resorption (synovial DOH pro and NBH levels); markers of lymphoid tissue activity (synovial and plasma beta 2m levels). Higher amounts of NDOH pro in RA synovial fluid are compatible with the hypothesis of a local connective tissue production as suggested by Uitto et al. on basis of a higher protocollagen proline hydroxylase activity in RA synovial tissue. DOH pro and NBH do not differ significantly in synovial fluid from RA or gouty patients, but the correlations between these forms of OH pro and, respectively, synovial lymphocytes and polymorphonuclear leucocytes are indicative of different processes of connective tissue remodelling in the 2 conditions. Synovial beta 2m levels are a direct function of total synovial lymphocyte counts independently of the type of arthropathy being explored. The ratio of synovial to plasma beta 2m is systematically above unity in RA patients only.

Protocollagen proline hydroxylase in skin and muscle of some vertebrate species

Protocollagen proline hydroxylase activity was studied in skin and muscle of poikilothermic and homoiothermic animals. Lower PPH activity was found in the former than in the latter. Increased PPH activity was found in skin of young Gallus domesticus and Rattus norvegicus. In specialized skin under hormonal control, i.e. comb and wattles, increased activity in relation to sexual maturation was shown. The PPH activity in muscles of poikilotherms was lower than in those of homoiotherms. Some of the muscles had a considerable enzymatic activity. A decreasing PPH activity was noticed in birds and mammals with age. Heart muscle had more hydroxylating activity than skeletal muscle.

Effects Of Some Connective-Tissue Active Drugs On Protocollagen Proline Hydroxylase Activity

The effect on protocollagen proline hydroxylase of drugs reported to be capable of inducing the lupus erythematosus syndrome and in some way acting on connective tissue and inflammation was studied in an in vitro system for the hydroxylation of 14C-Pro-labelled protocollagen. Some derivatives were also studied. It was found that benzoic acid and phenothiazine derivatives, hydroxyphenols, (+)catechin, beta-amino propionitrile, certain B vitamins, dihydrazinophthalazine, cysteine and dimethylcysteine were inhibitors of PPH. The chelation of Fe2+ions by the compounds mentioned is suggested to be essential.

Collagen changes in rheumatoid synovium

Thesynovium isnormally asingle ordouble layer ofcells withathin weaveofunderlying connective tissue. Inrheumatoid disease thesynovium becomes swollen andcongested, forming villous processes. Uitto etal.havedemonstrated increased protocollagen proline hydroxylase activity (1970b) anda concomitant increase incollagen synthesis (1972) in rheumatoid ascompared withnormalsynovium. Probably this inflammatory proliferation ofsynovial collagen formsanintegral partofthedisease process. We thought itimportant, therefore, to investigate thestructure ofcollagen innormal synovium andthechanges whichoccur inrheumatoiddisease. Thispaperdescribes ourfindings concerning theproportions oftypeIandtypem collagen innormal andrheumatoid synovium and thenature oftheintermolecular cross-links. Finally, theeffect ofD-penicillamine onrheumatoid synovial collagen will bediscussed. Investigations andresults We collected rheumatoid material from10patients atsynovectomy anddissected normalsynovium fromthekneesofninecadavers ofagesranging between three days and96years. Normal synovium wasdissected mostly fromtheinfrapatellar fatpad. We also tooksamples fromthesuprapatellar bursa

Stimulation of protocollagen proline hydroxylase activity by nucleoside triphosphates

Abstract Activity of purified protocollagen proline hydroxylase was enhanced several fold by addition of nucleoside triphosphates (3 mM) to the assay medium, but nucleoside mono-and diphosphates were almost inactive. Pyrimidine nucleotides were less effective compared with purine nucleotides, among which GTP was the most effective. dATP and ATP analogues such as adenosine 5′-(β,γ-imino) triphosphate (AMP-PNP), adenosine 5′-(β,γ-methylene) triphosphate (AMP-PCP), etc. were inactive. ATP or GTP showed no additive effect on enzyme activity stimulated by dithiothreitol or bovine serum albumin.

Effects of topical glucocorticoid medication on collagen biosynthesis in the dental pulp

The aim of the study was to observe pulpal collagen synthesis in response to trauma and to glucocorticoid medication. The material consisted of 290 rabbit pulps and 76 human premolar pulps. Collagen synthesis was determined by incubating whole pulps in a medium containing [14C]proline, and measuring the formation of [14C]hydroxyproline. The effect of glucocorticoids was studied in vitro using rabbit pulps. Hydrocortisone and dexamethasone inhibited collagen synthesis, whereas prednisolone had no marked effect. Hydrocortisone was found to inhibit the synthesis of [14C]hydroxyproline in neutral salt soluble and insoluble non-dialyzable collagen fractions. [14C]hydroxyproline in the dialyzable fraction was increased, suggesting that hydrocortisone increased collagen degradation. In the human material, premolar pulps were experimentally exposed and then medicated with capping agents. The contralateral teeth were exposed and capped with other capping materials, in some cases they were left as intact controls. The exposure led to an increase in the collagen synthesis as indicated by increased [14C]hydroxyproline formation and elevated protocollagen proline hydroxylase activity in the pulp. This enzyme activity was suppressed in pulps capped with a glucocorticoid paste. In addition, the collagen synthesis rate was lower in pulps treated with another glucocorticoid containing compound, when compared to pulps capped with a calcium hydroxide preparation.

Effects of glucocorticoid on connective tissue of aorta and skin in rabbits. Biochemical studies on collagen, glycosaminoglycans, DNA and RNA.

ABSTRACT Male rabbits were injected daily for 14 days with prednisone 2 mg/kg. The content and metabolism of collagen and glycosaminoglycans, the content of RNA, DNA, fat and water and the 125I-albumin permeability were investigated in the skin and in the intima-media layer of the aorta. A saline injected group served as control. Since prednisone induced a decrease in body weight, a starvation group with a similar weight loss was included. Following injections of prednisone the protocollagen proline hydroxylase activity, 0.45 m NaCl soluble hydroxyproline, total hydroxyproline, dialysable 14C-hydroxyproline and non-dialysable 14C-hydroxyproline fractions were reduced in the skin. In the aorta, only the total 14C-hydroxyproline synthesis was decreased. Furthermore there was a decrease in 14C-proline incorporation and the alpha-amino nitrogen content in the skin as well as a decrease in the RNA content in the skin and the aorta. It is concluded that prednisone acts mainly anti-anabolically on the metabolism of collagen and that this effect is part of a general inhibition of the protein synthesis. The sensitivity of the connective tissue of different organs to prednisone varies. Following prednisone treatment the 35S-sulphate incorporation into glycosaminoglycans was decreased in the skin as well as in aorta. However, this did neither affect the concentration nor the total amount of the different glycosaminoglycans – except for a decrease in chondroitin-4/6-sulphate in aorta. These findings probably indicate that prednisone inhibits the synthesis as well as the catabolism of the glycosaminoglycans. The skin and aorta reacted almost similarly. Prednisone raised the tissue to serum ratio of 125I-albumin. This may reflect changes in filtration and/or diffusion rates of albumin through the aortic endothelium and media. Finally, a decrease in body weight following starvation influenced the metabolism of collagen somewhat similar to the effect of prednisone, whereas the effects on the glycosaminoglycans differed.

Protocollagen proline hydroxylase activity in work-induced hypertrophy of rat muscle.

ARTICLESProtocollagen proline hydroxylase activity in work-induced hypertrophy of rat muscleH Turto, S Lindy, and J HalmeH Turto, S Lindy, and J HalmePublished Online:01 Jan 1974https://doi.org/10.1152/ajplegacy.1974.226.1.63MoreSectionsPDF (736 KB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat Previous Back to Top Next Download PDF FiguresReferencesRelatedInformation Cited ByMechanische Belastung und Bindegewebe17 August 2017Regulation of synthesis of fibrillar collagens in rat skeletal muscle during immobilization in shortened and lengthened positions20 December 2001 | Acta Physiologica Scandinavica, Vol. 172, No. 2Effects of prolonged exposure to and physical training in hypobaric conditions on skeletal muscle morphology and metabolic enzymes in ratsPflügers Archiv - European Journal of Physiology, Vol. 432, No. 1Effects of streptozotocin-induced diabetes, physical training and their combination on collagen biosynthesis in rat skeletal muscleActa Physiologica Scandinavica, Vol. 155, No. 1Extracellular matrix arrangement in the papillary muscles of the adult rat heart. Alterations after doxorubicin administration and experimental hypertensionBasic Research in Cardiology, Vol. 89, No. 4Quadriceps Exercises for Osteo-arthritis of the Knee — A Single Case Study Comparing Short-term versus Long-term Training EffectsPhysiotherapy, Vol. 80, No. 4Collagens in the injured porcine intervertebral disc18 February 2005 | Journal of Orthopaedic Research, Vol. 12, No. 1Enhanced prolylhydroxylase activity in the posterior annulus fibrosus of canine intervertebral discs following long-term running exerciseEuropean Spine Journal, Vol. 2, No. 3The effects of anabolic steroids on collagen synthesis in rat skeletal muscle and tendon23 April 2016 | The American Journal of Sports Medicine, Vol. 20, No. 3Alterations in collagen cross-linking impair myocardial contractility in the mouse heart.Circulation Research, Vol. 65, No. 6Cardiac interstitium in health and disease: The fibrillar collagen networkJournal of the American College of Cardiology, Vol. 13, No. 7Resistance Training: Physiological Responses and Adaptations (Part 2 of 4)12 July 2016 | The Physician and Sportsmedicine, Vol. 16, No. 4Collagen remodeling of the pressure-overloaded, hypertrophied nonhuman primate myocardium.Circulation Research, Vol. 62, No. 4The Passive Mechanical Properties of Muscle and Their Adaptations to Altered Patterns of UseAustralian Journal of Physiotherapy, Vol. 34, No. 3Collagen metabolism of mouse skeletal muscle during the repair of exercise injuriesPfl�gers Archiv European Journal of Physiology, Vol. 407, No. 1Collagen of slow twitch and fast twitch muscle fibres in different types of rat skeletal muscleEuropean Journal of Applied Physiology and Occupational Physiology, Vol. 52, No. 2Skeletal Muscle Adaptability: Significance for Metabolism and Performance1 January 2011Increased activities of prolyl 4-hydroxylase and galactosylhydroxylysyl glucosyltransferase, enzymes of collagen biosynthesis, in skeletal muscle of endurance-trained micePfl�gers Archiv European Journal of Physiology, Vol. 399, No. 4Biomechanics of the Musculotendinous Unit: Relation to Athletic Performance and InjuryClinics in Sports Medicine, Vol. 2, No. 1Muscle ultrastructural characteristics of elite powerlifters and bodybuildersEuropean Journal of Applied Physiology and Occupational Physiology, Vol. 48, No. 1Relationship between Muscle Hypertrophy and the Composition of Skeletal MusclesConnective tissue changes in surgically overloaded muscleCell And Tissue Research, Vol. 221, No. 2Connective tissue of “fast” and “slow” skeletal muscle in rats…effects of endurance trainingActa Physiologica Scandinavica, Vol. 108, No. 2Protein and collagen synthesis in rat diaphragm muscle incubated in vitro: The effect of alterations in tension produced by electrical or mechanical meansInternational Journal of Biochemistry, Vol. 11, No. 1Effects of physical training on metabolism of connective tissues in young miceActa Physiologica Scandinavica, Vol. 108, No. 1Effects of the inhibition of DNA synthesis on hypertrophying skeletal muscle.Journal of Biological Chemistry, Vol. 253, No. 9Oxidative metabolism of hypertrophic skeletal muscle in the ratExperimental Neurology, Vol. 48, No. 2Effect of Prolonged Physical Training on the Development of Connective Tissues in Growing MiceEnzyme activities in muscle and connective tissue of M. vastus lateralis in habitually training and sedentary 33 to 70-year-old menEuropean Journal of Applied Physiology and Occupational Physiology, Vol. 34, No. 1 More from this issue > Volume 226Issue 1January 1974Pages 63-65 Copyright & PermissionsCopyright © 1974 by American Physiological Societyhttps://doi.org/10.1152/ajplegacy.1974.226.1.63PubMed4358853History Published online 1 January 1974 Published in print 1 January 1974 Metrics

Effect of triiodothyronine in the metabolism of collagen in cultured embryonic bones.

The metabolism of collagen was studied with 14C-proline in organ cultures of embryonic ulnas. The addition of 0.01 ng-1.0 μg/ml of triiodothyronine caused a decrease in the incorporation of 14C-proline into protein. The amount of 14C-hydroxyproline was reduced even more, and a decrease was observed in the ratio of 14C-hydroxyproline to total 14C. In experiments with purified protocollagen proline hydroxylase, triiodothyronine did not inhibit the hydroxylation of protocollagen labeled with 14Cproline. Nor did cultivation of the ulnas with triiodothyronine have any effect on the total protocollagen proline hydroxylase activity in the bones. The degradative release of peptide-bound 14Chydroxyproline from the ulnas into the culture medium was increased in the presence of triiodothyronine, but there was no consistent increase in the release of free 14C-hydroxyproline. The results suggest that in our experimental conditions triiodothyronine caused a decrease in the accumulation of collagen in the ulnas. This ef...

Protocollagen proline hydroxylase in isolated rat liver cells

1. 1. Liver parenchymal cells from rats were dispersed by collagenase and hyaluronidase digestion, and by application of the bovine serum albumin flotation method. The viable cells, which were not stained with trypan blue, could be isolated with more than 95% viability. Contamination by non-parenchymal cells was less than 4% 2. 2. The endothelial cell fraction from rat liver was isolated by a combination of pronase digestion and bovine serum albumin flotation methods. Contamination by parenchymal cells was less than 0.4% and viability estimated with trypan blue was more than 99.8%. 3. 3. Protocollagen proline hydroxylase activity was assayed in both the parenchymal cell and the endothelial cell fractions. The parenchymal cells showed hydroxylase activity 100 times higher than the endothelial cells in terms of the number of cells. Trypan blue-stained parenchymal cells had about a quarter of the hydroxylase activity of viable parenchymal cells.

Protocollagen proline hydroxylase activity in rat heart during experimental cardiac hypertrophy.

Cardiac hypertrophy was produced in rats by constricting the abdominal aorta subdiaphragmatically. The weight of the left ventricle was significantly elevated 2 days after aortic constriction and was 19% higher than in shamoperated animals at 11 days. The activity of protocollagen proline hydroxylase (PPH), an enzyme participating in collagen biosynthesis, and the hydroxyproline content of the heart muscle were determined. PPH activity was increased at 2 days after aortic constriction and declined thereafter, being still above the control level at 11 days. The hydroxyproline content of the left ventricle was significantly increased at 11 days in hypertrophied heart muscle compared to controls. The present results suggest that in cardiac hypertrophy an early connective tissue activation occurs. Later, this leads to connective tissue accumulation. The increased collagen content may give support to the cardiac muscle contracting against systolic overload.

Injury and repair in arterial tissue in the rabbit. Analysis of DNA, RNA, hydroxyproline, and lactate dehydrogenase in experimental arteriosclerosis.

The aortic wall in rabbits was injured by pulling an inflated balloon catheter through the aorta. This was followed by an increase of the RNA-DNA ratio in the aortic wall 3 days after the injury. At the same time, the cathodic lactate dehydrogenase (LDH) isoenzymes composed of M subunits increased. The changes in LDH may reflect tissue hypoxia at the early phase after the injury. At 6 days protocollagen proline hydroxylase activity was increased, suggesting increased capacity for collagen formation in the injured aorta. Two weeks after injury the total content of hydroxyproline and DNA were increased, indicating connective tissue proliferation and hyperplasia in the injured aorta. The biochemical changes were interpreted as a repair response of the arterial connective tissue to injury. Gross arteriosclerosis of the aorta developed simultaneously with the biochemical alterations.

Protocollagen proline hydroxylase and collagen synthesis in developing eggs of Ascaris lumbricoides

1.1. Protocollagen proline hydroxylase (PPH) activity in developing eggs of Ascaris lumbricoides first appeared in the late blastula-early gastrula stages, was most active at or near the molting stage and disappeared in infective eggs. PPH activity was closely correlated with the amount of soluble collagen.2.2. Little hydroxyproline was present in gelatin extracted from the insoluble proteins of one-celled eggs. It increased forty-hold during development, whereas the gelatin increased only three-fold.3.3. The apparent Michaelis constant (Kma) of egg PPH was strongly affected by temperature, being minimal at 30°C. That of PPH from adult A. lumbricoides muscle was less strongly affected, with a minimum at 38°C.4.4. Oxygen concentrations exceeding 5 vol. per cent inhibited muscle but not egg PPH. Catalase did not relieve the inhibition.5.5. Egg and muscle PPH gave electrophoretically distinct peaks on acrylamide gel columns.6.6. The protocollagen proline hydroxylases of eggs and muscle seem to be isoenzymes which are adapted to the developmental physiology of A. lumbricoides.

Effect of a single injection of betamethasone disodium phosphate on the synthesis of collagen and noncollagen protein of carrageenin granuloma in rats

The effect of a single injection of betamethasone disodium phosphate on the incorporation of [ 3H]proline into collagen and noncollagen protein of rat carrageenin granuloma was studied. In all series of experiments, both [ 3H]proline and the steroid were given intravenously and the animals were killed at 30 min after the labeled proline injection in order to investigate precisely the time course of the drug action. The incorporation was not influenced at all by the steroid when it was given 30 min before the injection of [ 3H]proline. The inhibitory effect of the steroid, for both the collagen and the noncollagen protein, became significant at 1 hr after its intravenous administration, increasing progressively with the passage of time. Inhibition of the labeled proline incorporation into collagen hydroxyproline was significantly greater than that into noncollagen protein. Neither accumulation of protocollagen nor any change of protocollagen proline hydroxylase activity was demonstrated in the granuloma obtained from the rats treated with the steroid. The results show that the steroid does not inhibit the process of hydroxylation of protocollagen, suggesting indirectly the inhibition of synthesis of protocollagen or of [ 3H]proline transport through cell membrane, or of both.

Increased Protocollagen Proline Hydroxylase Activity in Sclerodermatous Skin

Protocollagen proline hydroxylase (PPH) activity in skin was determined in vitro by radioassay employing a tritium-labeled L-proline-peptide substrate. Average PPH activity was significantly higher in biopsy specimens of both forearm and parasacral skin from 20 patients with scleroderma than in specimens from ten normal subjects and 17 patients with other diseases. These findings offer evidence that collagen synthesis is increased in the skin in scleroderma and suggest that assay of PPH activity in skin may be a useful tool for the diagnosis of this disease.

Increased protocollagen proline hydroxylase activity in sclerodermatous skin

Increased protocollagen proline hydroxylase activity in sclerodermatous skin

Collagen metabolism in granulating wounds

A technique for studying small granulating wounds in man and laboratory animals is described. The wounds are produced and later excised using standard skin punches. Using this tissue we have determined the profile of a number of biochemical steps in collagen biosynthesis occurring in the early stages of wound healing. The most useful indicator of collagen synthesis was measurement of protocollagen proline hydroxylase (PPH) activity the enzyme which hydroxylates peptide-bound proline. In rat skin PPH-activity reaches a maximum 5 days after wounding. During starvation, the level of PPH-activity falls within a few days as does the soluble collagen content indicating decreased collagen synthesis. Systemic cortisone administration decreases the PPH level in wounds, and the effect can be reversed by adding systemic vitamin A (which has no effect by itself). Preliminary studies in humans indicate that similar elevations in PPH-activity occur in 5-day-old wounds. Patients with scleroderma have higher baseline levels of PPH-activity in their skin but show a normal response of PPH-activity to wounding.

Protocollagen Proline Hydroxylase Activity in Blood.

Excerpt We have developed new techniques to measure protocollagen proline hydroxylase (PPH) activity in human blood; PPH is the enzyme that hydroxylates proline during collagen synthesis. To determ...

Increased protocollagen proline hydroxylase activity in synovial tissue in rheumatoid arthritis.

Abstract Protocollagen proline hydroxylase activity was assayed in biopsy specimens of human synovial tissue. The enzyme required ascorbate, α-ketoglutarate, ferrous iron and molecular oxygen for its activity. The activity, determined from synovial tissue of 18 patients with rheumatoid arthritis, was increased as compared with values obtained from control patients.

Protocollagen proline hydroxylase activity in experimental pulmonary fibrosis of rats.

Abstract The activity of the enzyme protocollagen proline hydroxylase which synthesizes the hydroxyproline in collagen by hydroxylating proline in protocollagen was determined in rat lung tissue. About 80 per cent of the enzyme activity was recovered in the 100,000 × g supernatant fraction of the lung homogenate, and the enzyme required ascorbic acid, α-ketoglutarate, and ferrous iron for its activity. When ferrogluconate (100 mg. per kilogram per day) was injected into rats for 10 days, the enzyme activity increased into 134 per cent of the control value. Injections of ascorbate (100 mg. per kilogram per day) or cortisone (20 mg. per kilogram per day), or administration of d-penicillamine (200 mg. per kilogram per day) in drinking water did not change the enzyme activity. The protocollagen proline hydroxylase activity was determined in the lung tissue of rats with experimental silicosis induced by an intratracheal injection of silica dust. The enzyme activity was increased at 5 weeks in silicotic rats and reached a maximum, 259 per cent of the control value, at 16 weeks after the induction of silicosis. The enzyme activity was increased before fibrosis could be demonstrated by means of histological staining or chemical hydroxyproline analyses.