Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.4.22.6 (chymopapain)
407 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

To attempt to understand the etiology of failures of chemonucleolysis, biochemical analyses were performed on intervertebral disk material to determine if the enzyme had actually digested the nucleus pulposus proteoglycans. This information was then correlated with the clinical laboratory data to see if a pattern evolved for the failures. Nine chymopapain treated disks, 6 untreated herniated disks and 6 lumbar disks from scoliotic patients were obtained at surgery. The results indicated that 6 out of 9 patients treated with chymopapain had a marked reduction in the proteoglycan (hexosamine) content of their disk compared to the untreated controls. There was a significant inverse correlation of intrinsic lysosomal enzymes and hexosamine content in those cases where the chymopapain failed to destroy the proteoglycan. The other 3 patients, however, had hexosamine levels virtually identical to those disks not treated with chymopapain. The clinical evaluation, consisting of preoperative myelograms, diskograms, the surgeon's observations at laminectomy and evaluation of the postoperative regimen did not explain the failures. This study suggests that the chymopapain failures are not the result of inactivity of the enzyme or failure to digest the nuclear material in at least 6 of the 9 cases. However, there were 3 patients where either the enzyme was not reaching the nuclear material or it was inactive.
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PMID:Combined biochemical and clinical investigation of chemonucleolysis failures. 59 99

Proteoglycans were extracted from nuclease-digested sonicates of 10(9) rat basophilic leukemia (RBL-1) cells by the addition of 0.1% Zwittergent 3-12 and 4 M guanidine hydrochloride and were purified by sequential CsCl density gradient ultracentrifugation, DE52 ion exchange chromatography, and Sepharose CL-6B gel filtration chromatography under dissociative conditions. Between 0.3 and 0.8 mg of purified proteoglycan was obtained from approximately 1 g initial dry weight of cells with a purification of 200-800-fold. The purified proteoglycans had a hydrodynamic size range of Mr 100,000-150,000 and were resistant to degradation by a molar excess of trypsin, alpha-chymotrypsin, Pronase, papain, chymopapain, collagenase, and elastase. Amino acid analysis of the peptide core revealed a preponderance of Gly (35.4%), Ser (22.5%), and Ala (9.5%). Approximately 70% of the glycosaminoglycan side chains of RBL-1 proteoglycans were digested by chondroitinase ABC and 27% were hydrolyzed by treatment with nitrous acid. Sephadex G-200 chromatography of glycosaminoglycans liberated from the intact molecule by beta-elimination demonstrated that both the nitrous acid-resistant (chondroitin sulfate) and the chondroitinase ABC-resistant (heparin/heparan sulfate) glycosaminoglycans were of approximately Mr 12,000. Analysis of the chondroitin sulfate disaccharides in different preparations by amino-cyano high performance liquid chromatography revealed that 9-29% were the unusual disulfated disaccharide chondroitin sulfate di-B (IdUA-2-SO4----GalNAc-4-SO4); the remainder were the monosulfated disaccharide GlcUA----GalNAc-4-SO4. Subpopulations of proteoglycans in one preparation were separated by anion exchange high performance liquid chromatography and were found to contain chondroitin sulfate glycosaminoglycans whose disulfated disaccharides ranged from 9-49%. However, no segregation of subpopulations without both chondroitin sulfate di-B and heparin/heparan sulfate glycosaminoglycans was achieved, suggesting that RBL-1 proteoglycans might be hybrids containing both classes of glycosaminoglycans. Sepharose CL-6B chromatography of RBL-1 proteoglycans digested with chondroitinase ABC revealed that less than 7% of the molecules in the digest chromatographed with the hydrodynamic size of undigested proteoglycans, suggesting that at most 7% of the proteoglycans lack chondroitin sulfate glycosaminoglycans.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Purification and characterization of protease-resistant secretory granule proteoglycans containing chondroitin sulfate di-B and heparin-like glycosaminoglycans from rat basophilic leukemia cells. 241 30

Chymopapain (Discase) was injected at a dose of 0.125 nanokatal unit into the intervertebral discs of rabbits, and sequential changes in the metabolism of water, proteoglycan, collagen, and noncollagenous protein were investigated separately in the nucleus pulposus, anterior, and posterior anulus fibrosus. One week after chymopapain injection, the water and proteoglycan content was lower in all of the fractionated tissues of the anterior and posterior anulus and nucleus pulposus of the discs than in the control discs. In the anterior and posterior anulus, the proteoglycan content recovered after 12 weeks, but there was no recovery in the nucleus pulposus. The collagen content continued to increase up to the 12th week in the nucleus pulposus, while the noncollagenous protein content decreased in all tissue fractions after 1 week. In the anterior and posterior anulus, the content of noncollagenous protein recovered after 3 to 6 weeks, but there was no recovery in the nucleus pulposus. The lysine incorporation in collagen and noncollagenous protein was inhibited in all tissue fractions after 12 weeks, suggesting a decrease in synthetic activity. The intradiscal pressure calculated from proteoglycan hydration at 1 to 6 weeks after chymopapain injection showed a marked decrease to 0.8 to 0.9 atm, but it recovered to 1.6 atm after 12 weeks.
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PMID:Water, fixed charge density, protein contents, and lysine incorporation into protein in chymopapain-digested intervertebral disc of rabbit. 251 52

Changes on facet joint articular cartilage biology after chymopapain-induced disc height loss were determined in adult mongrel dogs. Disc height decreased to 50% at 2 weeks and returned to 80% at 6 months. The biochemistry and histology of the involved-level facet joints were followed at 2 weeks, 6 weeks, 3 months and 6 months after the injection. X-ray study showed a narrowing of the joint space and overlap of the facet joints at 6 weeks and at 3 months. Some return toward normal appearance was noted at 6 months. There was a loss of Safranin-O stain at 6 weeks, which persisted at 3 months, although some cells showed increased Safranin-O staining in the surrounding matrix. This was further improved at 6 months. The water content was unchanged early, but had decreased significantly at 6 weeks. The hexuronic acid content, already dropping at 2 weeks, was significantly lower at 6 weeks, at 3 months, and at 6 months. However, by 6 months, it had increased, compared with the 3-month value. Synthesis of proteoglycan was depressed only at 6 weeks. Similar changes were found in the facet cartilage in the joints above and below the last injected disc level. Results of this study would suggest that there is a cause-and-effect relationship between disc pathology and facet pathology and this can affect adjacent disc facet-joint biology. The initial facet lesion described appears to be potentially reversible, but a long-term disc height decrease might be expected to cause irreversible osteoarthritic-like changes in the adjacent facet joint.
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PMID:Facet joint changes after chemonucleolysis-induced disc space narrowing. 295 Jun

Following injection of chymopapain into a single knee joint in rabbits, serum keratan sulfate levels rose sharply and remained elevated for at least 48 hours before returning to preinjection levels. These changes were accompanied by depletion of proteoglycans from articular cartilage in the injected joint. We conclude that serum keratan sulfate levels rise predictably following acute loss of proteoglycan from a single joint.
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PMID:Increase in levels of serum keratan sulfate following cartilage proteoglycan degradation in the rabbit knee joint. 296 77

Chymopapain (1 mg) was injected into each of four-lumbar intervertebral discs of adult mongrel dogs. As expected, at 2 weeks, all injected discs exhibited marked loss of height (mean: 50% of original height) indicative of severe proteoglycan depletion. The appearance of keratan sulfate-bearing fragments in plasma was monitored by an ELISA-inhibition assay which uses a monoclonal antibody (1/20/5-D-4) specific for an epitope present only in the longest keratan sulfate chains. Levels of plasma keratan sulfate rose within 30 minutes and reached a maximum between 24 and 72 hours later. Levels then declined progressively but were still elevated at 2 weeks postinjection. Keratan sulfate-bearing fragments in plasma were purified by ion exchange chromatography on DEAE-Sephacryl and fractionated by sieve chromatography on Sepharose CL-6B. These plasma keratan sulfate-bearing fragments were found to be similar in size to keratan sulfate-bearing fragments generated by chymopapain digestion of dog nucleus pulposus proteoglycans, but slightly larger than single keratan sulfate chains obtained by alkaline borohydride treatment of dog nucleus pulposus proteoglycans. The results of this study show that measurements of blood levels of keratan sulfate could prove useful in monitoring effective degradation of disc proteoglycans in chemonucleolysis in man and help discriminate between ineffective enzyme placement, and alternative mechanisms of treatment failure.
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PMID:Levels of keratan sulfate-bearing fragments rise predictably following chemonucleolysis of dog intervertebral discs with chymopapain. 297 75

To investigate the histopathological changes of the intervertebral disc after chemonucleolysis, experiments were performed on 21 monkeys. After the injection of chymopapain, animals were sacrificed at definite intervals up to 1 year. Histopathological studies on these specimens revealed disappearance of proteoglycan from the nucleus pulposus, inner annulus fibrosus and cartilaginous endplates as early as 1 day after injection. At 4 weeks, regeneration of proteoglycan was indicated by the recovery of positive Safranin-O (S-O) staining in the area of the nucleus pulposus. After 24 weeks, the entire intervertebral disc showed uniform S-O staining indicating further regeneration of proteoglycan. The matrix of the reformed nucleus pulposus contained increased amount of fibrous elements compared to the controls. These results indicate proteoglycan regeneration by chondrocytes after chemonucleolysis. The reformed nucleus was histopathologically different from the control.
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PMID:Chemonucleolysis--an experimental histopathological study. 313 15

The effects of chymopapain treatment of the canine intervertebral disc were studied in vivo by monitoring proteoglycan in the nucleus pulposus. Analysis of proteoglycan was carried out by Sepharose CL-4B (Pharmacia Fine Chemicals AB, Stockholm, Sweden) chromatography and electrophoresis. The proteoglycan was degraded to glycosaminoglycans within 1 week after chymopapain treatment. Two weeks later, a proteoglycan smaller than the original appeared in the nucleus pulposus. At 8 weeks after injection, the amount of the newly synthesized proteoglycan, similar in molecular weight to the original, had recovered to about half that of the original, although the new proteoglycan fraction was rich in hyaluronic acid. It was concluded that, following chemonucleolysis with chymopapain, the water-binding capacity of the nucleus pulposus recovered, but that the regenerated nucleus pulposus differed biochemically from the original.
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PMID:Degradation and biosynthesis of proteoglycans in the nucleus pulposus of canine intervertebral disc after chymopapain treatment. 320 93

Seventeen patients with intractable sciatica due to prolapse of a lumbar disc, treated by intradiscal injection of chymopapain (chemonucleolysis) were studied. Analysis of serial 24 hour urine collections showed a significant increase in urinary glycosaminoglycan after chemonucleolysis. This was not detected in four patients undergoing routine discography. Enzymic analysis of urinary glycosaminoglycan after chemonucleolysis suggested that the increase in levels was largely due to an increase in the amounts of chondroitin sulphate present, probably resulting from proteoglycan breakdown in the intervertebral disc. Eight of the patients treated by chemonucleolysis underwent serial computed tomography (CT). One month after the injection the only change seen was a loss of definition of the disc prolapse, which could be interpreted as a loss of turgidity in the disc as a result of proteoglycan breakdown by chymopapain. By six months the CT of those patients whose symptoms had improved showed that the degree of disc prolapse was usually less marked and the disc margin more clearly defined, suggesting that by this stage anatomical remodelling had occurred.
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PMID:Mechanism of action of intradiscal chymopapain in the treatment of sciatica: a clinical, biochemical, and radiological study. 372 72

The three proteinases present in papaya latex: papain (EC 3.4.22.2) chymopapain and papaya proteinase III (EC 3.4.22.6), were standardized by active-site titration, and compared in proteolytic activity against azocasein, serum albumin and cartilage proteoglycan. The activities were all of the same order, although there were differences in pH dependence. SDS-polyacrylamide gel electrophoresis of the early products of digestion of albumin and phosphorylase a showed very similar patterns for the three papaya proteinases. Kinetic parameters for hydrolysis of benzyloxycarbonyl-phenylalanyl-arginyl-7(4-methyl)coumarylamide were determined for the three enzymes. Values for kcat/Km varied only within a factor of 2, but the individual constants were much higher for papain than for chymopapain and papaya proteinase III. In contrast to the results obtained with the synthetic substrate, the kinetic parameters for the initial hydrolysis of succinyl-albumin were very similar for the three papaya proteinases. This was consistent with their similar proteolytic activities in other assays.
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PMID:The proteolytic activities of chymopapain, papain, and papaya proteinase III. 391 69


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