Catheptic Activity Research Articles

EFFECT OF GAMMA RADIATION ON CHICKEN LIVER CATHEPTIC ACTIVITY AND RELEASE OF LYSOSOMAL CATHEPSIN D

Journal of Food ScienceVolume 40, Issue 1 p. 47-49 EFFECT OF GAMMA RADIATION ON CHICKEN LIVER CATHEPTIC ACTIVITY AND RELEASE OF LYSOSOMAL CATHEPSIN D MUMTAZ ALI, MUMTAZ ALI Dept. of Food Science, University of British Columbia, Vancouver, Canada V6T 1W5Search for more papers by this authorJ. F. RICHARDS, J. F. RICHARDS Dept. of Food Science, University of British Columbia, Vancouver, Canada V6T 1W5Search for more papers by this author MUMTAZ ALI, MUMTAZ ALI Dept. of Food Science, University of British Columbia, Vancouver, Canada V6T 1W5Search for more papers by this authorJ. F. RICHARDS, J. F. RICHARDS Dept. of Food Science, University of British Columbia, Vancouver, Canada V6T 1W5Search for more papers by this author First published: January 1975 https://doi.org/10.1111/j.1365-2621.1975.tb03732.xCitations: 7 One of the authors (Mumtaz Ali) was supported by the Canadian International Development Agency during the course of this study. The research reported herein was made possible by a contract with Atomic Energy of -Canada Ltd. and a National Research Council Operating Grant. The authors thank Dr. R.C. Whiting for reviewing the manuscript. AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Citing Literature Volume40, Issue1January 1975Pages 47-49 RelatedInformation

MUSCLE CATHEPSINS IN THREE SPECIES OF PACIFIC SOLE

The cathepsins in saline muscle extracts of three species of Pacific sole were investigated. A buffered hemoglobin solution (pH 3.0) served as substrate for the enzyme assay. No apparent change in catheptic activity in muscle homogenates of English sole was observed during 25 days of frozen (−26 °C) storage. Partial characterization of the cathepsins indicated a pH activity optimum of 3.0–3.5 for rex and petrale sole but a slightly higher PH optimum of 3.2–3.8 for English sole. The three species showed a temperature optimum of 45 °C for hemoglobin splitting. A wide variation in catheptic activity within the species was observed for rex sole while English and petrale sole showed more uniform catheptic activity. The mean activities for the three species varied significantly (P < 0.05). Results of sensory evaluation of muscles containing high and low levels of catheptic activity from the three species of sole did not support the assumption that cathepsins have a substantial influence upon the quality of marine food fish.

CATHEPTIC ENZYME ACTIVITY IN AGED COUNTRY‐STYLE HAMS AS INFLUENCED BY PRE‐CURING TREATMENT

Chemical composition and catheptic activity were determined in fresh chilled hams and four lots of cured hams subjected to different pre‐curing treatments. Hams were sampled after aging (storage) of 2, 4 and 6 months at 29 °C. Moisture decreased, while protein, salt and free amino acid concentrations increased with aging time of cured hams, but treatment effects were similar. Frozen, thawed cured hams aged for 2 months exhibited the highest specific enzyme activity but decreased in enzyme activity after 4 and 6 months of aging. Free amino acid concentration after aging was higher in hams frozen 1–3 yr (X̄= 2.3) at −29 °C than for those frozen 0 or 2 days.

FACTORS RESPONSIBLE FOR WHITE FILM FORMATION ON CUT SURFACES OF DRYCURED HAMS

White surface film that forms on cut surfaces of country‐style ham was identified as crystalline L‐tyrosine. Free L‐tyrosine averaged 287.3 μmoles/g (5.2% by weight) in the film and 6.1 μmoles/g in ham tissue from which film had been removed. It is concluded that the film forms because the concentration of free L‐tyrosine and certain other free amino acids in ham is greater than solubility levels; therefore, crystals form on the cut surface. The absence of surface film on knuckle muscles and film formation on other muscles of the same slice was attributed to higher NaCl (P < 0.05) of the former which decreased catheptic activity.

Effect of ATP on protein degradation in rat liver lysosomes.

The catheptic activity of rat liver lysosomes was found to be stimulated by the addition of ATP plus Mg ions. It is concluded that ATP facilitates the pinocytotic engulfment of proteins into the lysosomes and thereby stimulates proteolysis.

CATHEPTIC ACTIVITY OF FISH MUSCLE

Catheptic activity was found to be localized in the lysosomal fraction of skeletal muscle of winter flounder (Pseudopleuronectes americanus). The enzyme hydrolyzed hemoglobin, globin, bovine serum albumin and endogenous proteins. The hemoglobin splitting activity of the enzyme under different conditions was determined by gel electrophoresis. An optimum pH of 4.0 was found for both the hemoglobin breakdown and autolytic activity on the endogenous proteins. The enzyme was highly thermostable, dialysis had no effect on its activity, and the enzyme was inhibited by sodium chloride, cysteine, ATP, NAD and phenyl methyl sulfonyl fluoride. Its molecular weight was found to be about 32,000 using sucrose gradient centrifugation.

The limited proteolysis of bovine neurophysins by cathepsin D.

Abstract— The limited proteolysis of the bovine neurophysins at acid pH has been studied and the enzyme responsible has been characterized. Only 15 per cent of the catheptic activity in 4‐year‐old acetone‐dried posterior pituitary lobe powder is soluble at pH 4.0. Solubility increases as the age of the powder decreases and the cathepsin is completely soluble in the presence of 1% Triton X‐100. Acid proteinase activity in the neurohypophysis is not thiol activated and is inhibited by 3‐phenylpyruvic acid. Bovine serum albumin was degraded at only 1 per cent of the rate of haemoglobin but with the same pH optimum (3.7). On this basis the enzyme was identified as cathepsin D.Neurophysin‐I is degraded in two stages by cathepsin D; the first product (neurophysin‐I′) runs faster and the second product (neurophysin‐I″) runs slower than the native protein on starch‐gel electrophoresis at pH 8.1. Neurophysin‐II is also degraded in two stages; the first product has a higher electrophoretic mobility than the native protein and is identical in mobility with the faster‐running component of the so‐called neurophysin‐M of Hollenberg and Hope (1967b). Prolonged incubation with the cathepsin gives rise to a slower‐running component. Neurophysin‐C is not attacked by the acid proteinase. Neurophysin‐I′ and I″ have been isolated by ion‐exchange chromotography. They have the same N‐terminal amino acid (alanine) and C‐terminal sequence (Ala‐Phe‐Ser) as the native protein and both bind 8‐argininevasopressin. Neurophysin‐I′ is identical in amino acid composition with the native protein but neurophysin‐I″ has lost one leucine and two aspartic acid residues.Reduction, 14C‐alkylation and separation of the fragments by starch‐gel electrophoresis shows that the structural and functional integrity of neurophysin‐I″ is maintained by the disulphide bonds, even though a tripeptide has been split out of the interior of the molecule. The low molecular weight material produced by catheptic digestion of neurophysin‐I has been purified and shown to have a composition of one leucine and two aspartic acid residues. It is suggested that extensive in vivo proteolysis of neurophysin by lysosomal cathepsin, with consequent abolition of hormone‐binding ability, is unlikely.

Study of catheptic and acid phosphatase activities during development and metamorphosis ofDrosophila melanogaster.

Developmental stages ofDrosophila melanogaster cultured at 22 ± 1° C were collected and tested for catheptic activity and acid phosphatase activity.It was found that catheptic activity was absent in the egg as well as in the first and second larval instars. The activity first appears in the third instar larva and reaches its peak 24 to 48 hr after puparium formation. It then decreases, at first gradually and at pupal stage 9 (120 to 144 hr after puparium formation) abruptly, reaching zero level just before the emergence of the imago.The pattern of acid phosphatase activity in different developmental stages was found to be broadly similar to that of catheptic activity in the larval and pupal stages. However, the acid phosphatase activity was found to be exceptionally high in the egg in contrast to the catheptic activity.

Catheptic and Trypsin-inhibiting Activities in the Nutritional Muscular Dystrophy

From 50 days on, after vitamin E deficient diet was given to guinea pigs, typical symptoms of muscular dystrophy were observed. Muscle protein of these animals was fractionated into 3 fractions, sarcoplasmic fraction, Triton X–100 soluble fraction and myofibrillar fraction.Free catheptic activity (hemoglobin-splitting activity in the sarcoplasmic fraction), bound catheptic activity (the same in the Triton X–100 soluble fraction) and trypsin-inhibiting activity (in the sarcoplasm fraction) were determined in normal and dystrophic muscles.In dystrophic muscles, great increase in free and bound catheptic activities was observed.The increase of free catheptic activity was greater than that of bound catheptic activity.Trypsin-inhibiting activity was also increased in dystrophic muscles two- or three-fold. At the same time, decrease in the content of myofibrillar protein in dystrophic muscles was observed.These results were discussed in view of the degradation of protein in dystrophic muscles.

Comparative Study of the Muscle Catheptic Activity of Some Marine Species

The hemoglobin-splitting activity and the autolytic- (endogenous) catheptic activity of the white skeletal muscle of 13 Atlantic fish species were measured and found to be low relative to the catheptic activity of other tissues. On the other hand, catheptic activities of lobster and crab claw muscles were high. Lobster tail muscle showed little activity, a factor perhaps contributing to its tougher texture relative to the claw muscle. The individual variations between samples of the same species were great and the ranges of the species overlapped each other with no evident phylogenetic distinction. The hemoglobin-splitting activity was always higher and was always directly correlated with the autolytic activity for each individual sample. Using an urea-free hemoglobin substrate pH optima were found in the range 3.0–3.6. Urea shifted the pH optima upwards approximately 0.4 unit. Fish muscle cathepsins were found to have the same level of activity as chicken muscle.

Autolytic Breakdown of Rat Skeletal Muscle Proteins in the Alkaline pH Range

Properties of autolytic breakdown of rat skeletal muscle proteins in the alkaline pH range have been reported. The activity is almost exclusively localized in the myofibrillar fraction, but is not solubilized with Triton X-100. The activity is affected by the KCI concentration in the reaction mixture. In 0.6 M and the more concentrated KCI solutions, the maximum activity is attained. The optimum pH of the activity is in the range of pH 7.5∼9.5, and the optimum temperature is between 47∼57°C. This autolytic activity seems to be different from catheptic activity which shows its optimum pH in the acid pH range. Moreover, though more than half of the catheptic activity of rat skeletal muscle is recovered in the myofibrillar fraction, the catheptic activity in the myofibrillar fraction can be removed from the fraction by the extraction with dilute saline solution containing Triton X-100.

Autolytic Breakdown of Rat Skeletal Muscle Proteins in the Alkaline pH Range

Properties of autolytic breakdown of rat skeletal muscle proteins in the alkaline pH range have been reported. The activity is almost exclusively localized in the myofibrillar fraction, but is not solubilized with Triton X-100. The activity is affected by the KCI concentration in the reaction mixture. In 0.6 M and the more concentrated KCI solutions, the maximum activity is attained. The optimum pH of the activity is in the range of pH 7.5∼9.5, and the optimum temperature is between 47∼57°C.This autolytic activity seems to be different from catheptic activity which shows its optimum pH in the acid pH range. Moreover, though more than half of the catheptic activity of rat skeletal muscle is recovered in the myofibrillar fraction, the catheptic activity in the myofibrillar fraction can be removed from the fraction by the extraction with dilute saline solution containing Triton X-100.

Changes in catheptic activity in the enamel organ of the bovine tooth germ during development

Changes in catheptic activity in the enamel organ of the bovine tooth germ during development

Lysosomal enzyme activity in the interstitial fluid of solid mouse tumour transplants

This study embodies lysosomal enzyme assays on cell-free interstitial fluid from peripheral and central regions of various solid mouse tumour transplants. Comparison was made with the corresponding activities of normal plasma and intraperitoneal fluid from the same strains of mice. All enzymatic activities have been expressed per a volume basis of either 1 or 10μl of original tumour fluid. The characteristic and somewhat unexpected feature was that central tumour fluid presented, on an average, very large increases in acid nitrophenyl-phosphatase, β-glucuronidase, aryl sulphatase and total catheptic activity at pH 5·0. Evidence was further presented for the occurrence of cathepsin B activity following the addition of cysteine + EDTA. The dual origin of added enzymatic proteins both from tumour and host cells is discussed. Among the lysosomal hydrolases described the greatest physiological interest is attributed to the trypsin-like cathepsin B activity. This activity seems to be operative in the detachment of tumour cells, thus favouring invasion and the formation of metastasis.

Subcellular Distribution of Human Leukocytic Cathepsins

Abstract 1. Methods are presented for the disruption of human peripheral leukocytes by sonication and the separation of the released particulate matter by differential centrifugation. 2. The distribution of catheptic activity in the various fractions was investigated both on material obtained from mixed populations of leukocytes and on that from isolated lymphocytes and granulocytes. 3. All of the leukocytic fractions tested contained cathepsins active at pH 3.5 and pH 8.5. 4. At pH 3.5, the catheptic activity was rather evenly distributed among nuclear (I), and light (IV) and heavy (II) granular fractions, with practically no activity in the soluble fraction (V). 5. At pH 8.5, there was always more catheptic activity in the heavier (II) than the lighter (IV) granules, variable amounts in the nuclear fraction (I) and very little in the supernatant (V). 6. At pH 3.5, the lymphocytic granules showed considerably higher proteolytic specific activity than those from granulocytes. 7. At pH 8.5, all fractions except the heavy granules (II) had higher proteolytic specific activities in granulocytes than in lymphocytes. 8. At pH 3.5, and to a lesser extent at pH 8.5, the light (IV) and heavy (II) granules from chronic lymphatic leukemia cells contained much lower cathepic activity than those from normal lymphocytes. 9. Cathepsins in each subcellular fraction are very labile, losing as much as 80 per cent of their activity upon storage at 4 C. overnight in distilled water.

Catheptic activity in the developing rat brain.

Catheptic activity in the developing rat brain.

Effect of methycellulose on catabolic processes in normal and irradiated guinea-pig spleen.

Catheptic activity and metabolic nitrogen was measured in guineapigs treated with methylcellulose, which induced hypersplenism.

Vergleichende untersuchungen der sauren proteolytischen enzymaktivitäten von serum und punktionsflüssigkeiten entzündlicher, nichtentzündlicher und maligner genese

1. 1. The pH-activity curves of the proteolytic enzymes from sera and exudates withdrawn by puncture from inflammatory, non-inflammatory and malignant tissue were examined in the pH range 0.9–4.4. Casein was used as substrate, and a sensitive copper complex method was employed. 2. 2. The proteolytic activity in sera and in exudates showed great quantitative and qualitative differences. The pH-activity curves of sera had a plateau between pH 1.2 and 2.0, and a single peak, between pH 2.2 and 2.6. The curves pertaining to exudates showed up to 4 peaks of greatly variable height, lying within a narrow range. 3. 3. The activity curves of the exudates derived from the three groups showed only quantitative differences, while peak distribution and the general aspect of the curves were similar. Fluids and sera of malignant origin had a higher catheptic activity than those from the other two groups. 4. 4. The results are discussed in connection with the problem of inflammation. The course of the pH-activity curves led to the conclusion that qualitative changes in the pH range 3.7–4.0 are only caused by processes accompanied by extensive cellular breakdown. The qualitative differences between the curves for sera and exudates are interpreted as representing nonspecific signs of cell damage caused by the effusion. An attempt is made to correlate the individual activity peaks with known proteolytic enzyme systems.

Catheptic Activity of Chicken Muscle: Quantitative Assay of the Hemoglobin‐Splitting Catheptic Activity

SUMMARY— A quantitative method was developed to assay the hemoglobin‐splitting catheptic activity in chicken muscle. The catheptic activity of chicken breast muscle was found to be 34‐fold greater than previously reported.

Relationship between age and haemoglobin-splitting activity in chicken muscle.

CATHEPTIC activity towards proteins has been found in extracts of kidney, spleen, liver, lung, muscle, brain and erythrocytes1–5; therefore, the cathepsins may be considered to be part of the general enzymatic apparatus of animal cells. The action of the cathepsins is evident during the autolysis of animal tissues at acid pH. Recently, considerable interest has been shown in the catheptic activity of beef and chicken muscle because of their suggested role in meat tenderization, preservation and flavour6–10.