scholarly journals Mono- and poly-ADP-ribosylation of proteins in mouse kidney after castration and testosterone treatment

1981 ◽  
Vol 198 (1) ◽  
pp. 37-44 ◽  
Author(s):  
A Gartemann ◽  
R Bredehorst ◽  
K Wielckens ◽  
W H Strätling ◽  
H Hilz
Keyword(s):  
Enzyme Activity ◽  
Total Protein ◽  
Nuclear Protein ◽  
Mouse Kidney ◽  
Testosterone Treatment ◽  
Control Value ◽  
Protein Conjugates ◽  
Testosterone Substitution ◽  
Testosterone Administration ◽  
Adp Ribosyltransferase

Protein-bound mono(ADP-ribose) and poly(ADP-ribose) residues were determined in mouse kidney after castration and testosterone substitution. After these treatments, the mouse kidney undergoes significant alterations in the extent and pattern of transcription without changes in the amount of DNA and nuclear protein. The amount of mono(ADP-ribose)-protein conjugates (the hydroxylamine-sensitive and -resistant subfractions) decreased by 40% after castration, and returned to normal within 1 week after daily testosterone injections. Polymeric ADP-ribose residues, which amounted to less than 0.3% of the total protein-bound monomeric ADP-ribose, increased after castration and rapidly decreased on testosterone administration. The magnitude of these effects indicates that the decrease in mono(ADP-ribose) was not caused by a shift of monomeric residues into the polymer form. Nuclear ADP-ribosyltransferase activity showed a retarded decrease after castration, reaching 60% of the control value by day 20. After testosterone injections, enzyme activity rose to normal within 3-4 days. The amounts of the substrate NAD+ as well as of NAD+ + NADH also declined after castration, and rapidly returned to values slightly above normal when the androgen was substituted. The differential response of monomeric and polymeric ADP-ribose residues to castration and testosterone treatment suggests that the two modifications serve different functions.

Heterogenous expression of ornithine decarboxylase gene in the proximal tubule of the mouse kidney following testosterone treatment

1993 ◽  
Vol 100 (5) ◽  
pp. 325-330 ◽  
Author(s):  
Noriyuki Koibuchi ◽  
Shigeru Matsuzaki ◽  
Mikako Sakai ◽  
Hideki Ohtake ◽  
Sadao Yamaoka
Keyword(s):  
Proximal Tubule ◽  
Ornithine Decarboxylase ◽  
Mouse Kidney ◽  
Testosterone Treatment

scholarly journals Increase in alkaline phosphatase activity in calvaria cells cultured with diphosphonates

1979 ◽  
Vol 183 (1) ◽  
pp. 73-81 ◽  
Author(s):  
R Felix ◽  
H Fleisch
Keyword(s):  
Alkaline Phosphatase ◽  
Protein Synthesis ◽  
Enzyme Activity ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Heat Inactivation ◽  
High Concentration ◽  
Control Value ◽  
Km Value ◽  
Inhibitor Of Protein Synthesis

1. Dichloromethanediphosphonate and to a lesser degree 1-hydroxyethane-1,1-diphosphonate, two compounds characterized by a P-C-P bond, increased the alkaline phosphatase activity of cultured rat calvaria cells up to 30 times in a dose-dependent fashion. 2. Both diphosphonates also slightly inhibited the protein synthesis in these cells. 3. Thymidine, an inhibitor of cell division, did not inhibit the induction of the enzyme, indicating that the increase in enzyme activity was not due to the formation of a specific population of cells with high alkaline phosphatase activity. 4. The effect on alkaline phosphatase was suppressed by the addition of cycloheximide, an inhibitor of protein synthesis. 5. After subculturing the stimulated cells in medium without diphosphonates, the enzyme activity fell almost to the control value. 6. Bovine parathyrin diminished the enzyme activity of the control cells and the cells treated with dichloromethanediphosphonate; however, at high concentration the effect of parathyrin was greater on the diphosphonate-treated cells than on the control cells. 7. The electrophoretic behaviour, heat inactivation, inhibition by bromotetramisole or by phenylalanine, and the Km value of the induced enzyme were identical with that of the control enzyme.

Development and regression of exercise-induced cardiac hypertrophy in rats

1979 ◽  
Vol 236 (2) ◽  
pp. H268-H272 ◽  
Author(s):  
R. C. Hickson ◽  
G. T. Hammons ◽  
J. O. Holoszy
Keyword(s):  
Cytochrome C ◽  
Protein Content ◽  
Total Protein ◽  
Female Rats ◽  
Heart Weight ◽  
Total Protein Content ◽  
Mitochondrial Marker ◽  
Time Intervals ◽  
Control Value ◽  
Exercise Induced

Adult female rats were exercised by daily swimming. All the increase in heart weight induced by the exercise occurred within 14 days and averaged 30%. The half times of the increases in heart weight and total protein content were about 4.5 days, whereas that of cytochrome c, which was used as a mitochondrial marker, was 6.5 days. The total amounts of DNA and of hydroxyproline in the heart, which were used to evaluate the degree of connective tissue hyperplasia, increased only slightly (8% and 10%, respectively). Other animals were subjected to the same swimming program for 21 days. Groups of rats were killed at various time intervals after stopping exercise. Heart weight, total protein content, and total cytochrome c content decreased rapidly initially, with 60% of the total regression of hypertrophy occurring during the first week. Thereafter, heart weight fell more gradually toward the sedentary control value. The hydroxyproline content of the heart, which was increased 10%, did not decrease during the regression of the hypertrophy.

scholarly journals Immunoassay of muscle-specific creatine kinase with a monoclonal antibody and application to myogenesis and muscular dystrophy

1983 ◽  
Vol 213 (2) ◽  
pp. 417-425 ◽  
Author(s):  
G E Morris ◽  
L P Head
Keyword(s):  
Monoclonal Antibody ◽  
Creatine Kinase ◽  
Enzyme Activity ◽  
Total Protein ◽  
Plasma Concentrations ◽  
Thigh Muscle ◽  
Cell Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Activity Measurements

A competition e.l.i.s.a. (enzyme-linked immunosorbent assay) is described that enables direct measurement of the muscle-specific polypeptide of chick creatine kinase (M-CK) in extracts of differentiating muscle-cell cultures and in blood plasma samples, even in the presence of embryonic, or brain-type, creatine kinase. The characteristics of the assay can be considerably improved by the use of a monoclonal antibody, CK-ART, instead of rabbit antisera, and we offer an explanation for this in terms of heterogeneity of antibody affinities in polyclonal antisera. In addition to native enzyme, the assay will measure creatine kinase unfolded and inactivated by 8 M-urea treatment. During chick muscle differentiation in vitro, M-CK increased from 7.5% of the total creatine kinase at 24h to 76.0% at 143h, in good agreement with isoenzyme separation data. As a percentage of the total cell protein, M-CK increased by 156-340-fold over the same period and constituted 0.38-0.56% of the total protein in late cultures. E.l.i.s.a. measurements on 17-20-day embryonic thigh-muscle extracts, which contain almost exclusively M-CK, agree well with enzyme activity and radioimmunoassay. M-CK constituted 0.7-1.6% of the total protein in 17-19-day embryonic thigh muscle. Plasma M-CK concentrations in normal 2-8-week-old chickens were found to be in the range 0.5-0.9 micrograms/ml. Plasma concentrations of 32-56 micrograms/ml were found in 8-week-old dystrophic chickens by both e.l.i.s.a. and enzyme-activity measurements. The results suggest that inactive or unfolded forms of M-CK do not normally exist, in any significant amounts, in cell and tissue extracts or in freshly prepared samples of plasma.

Changes in protein during development of Triturus embryos

Development ◽  
1973 ◽  
Vol 30 (3) ◽  
pp. 647-659
Author(s):  
Hiroshi Imoh ◽  
Tsutomu Minamidani
Keyword(s):  
Protein Synthesis ◽  
Protein Content ◽  
Sodium Carbonate ◽  
Dna Content ◽  
Total Protein ◽  
Soluble Protein ◽  
Nuclear Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Tail Bud ◽  
Stage Of Development

The present paper reports basic data on DNA content, protein content, and protein synthesis in Triturus pyrrhogaster embryos during development from cleavage to the hatching stage. Except for measurements of DNA and total protein contents, embryos were labeled with sodium carbonate-14C for 10 h and fractionated into embryonic cell components, i.e. cytoplasmic mass, yolk and pigment granules, and nuclei, in a discontinuous density gradient of sucrose. The protein content and the radioactivity incorporated into protein were measured in each fraction. Those fractions combining protein soluble in buffer at pH 8·3 and in 0·25 N-HCl were further studied with polyacrylamide gel electrophoresis. In the newt embryo, four stages of active DNA increase were observed when cultured at constant temperature; they were gastrula, neurula, late tail-bud, and before-hatching stages. Total protein per embryo decreased from 3 to 2 mg during the development studied. The content of cytoplasmic soluble protein per embryo was low and constant throughout development. Synthesis of the fraction was observed at the earliest stage of development studied though the rate was not high and specific activity of the soluble protein increased during development. Qualitative changes in the newly synthesized protein were observed. With the yolk fraction, synthesis of protein, other than from probable contamination with the cytoplasmic fraction, was not detected and a detailed description was omitted. Changes were observed at two stages of development in the synthesis of nuclear protein soluble in buffer at pH 8·3, the first at gastrulation and the second at late tail-bud stage. The change at gastrulation seemed to be the start of syntheses of the nuclear soluble proteins, while quantitative enhancement rather than qualitative change was noticed at late tail-bud stage. Most of the nuclear protein soluble in 0·25 N-HCI was histone. The histone content increased in accordance with increase in the DNA content and the rate of DNA accumulation was accompanied by proportionate incorporation of radioactivity into histone. Among histone fractions, unique behaviour of the very lysine-rich histone was observed. The availability of [14C]sodium carbonate in rough estimations of protein synthesis in embryos and significance of the data obtained have been discussed.

scholarly journals PURIFICATION AND PHYSICAL PROPERTIES OF GROUP C STREPTOCOCCAL PHAGE-ASSOCIATED LYSIN

1971 ◽  
Vol 133 (5) ◽  
pp. 1105-1117 ◽  
Author(s):  
Vincent A. Fischetti ◽  
Emil C. Gotschlich ◽  
Alan W. Bernheimer
Keyword(s):  
Molecular Weight ◽  
Enzyme Activity ◽  
Physical Properties ◽  
Total Protein ◽  
Sedimentation Coefficient ◽  
Polyacrylamide Gel ◽  
Single Band ◽  
Purification Procedure ◽  
Sh Group ◽  
Stokes Radius

A purification procedure for the Group C phage-associated lysin is described utilizing tetrathionate to protect the enzyme's -SH group(s) from thiol-inactivating agents. A 652-fold purification has been accomplished yielding a solution in which the enzyme activity corresponds to essentially a single band on polyacrylamide gel which accounts for 70% of the total protein in the preparation. A molecular weight of 101,000 and frictional ratio of 1.526 was determined for the lysin utilizing experimentally determined values for its Stokes radius and sedimentation coefficient.

scholarly journals Expression of heme oxygenase-1 in the lung in chronic hypoxia

2000 ◽  
Vol 278 (4) ◽  
pp. L806-L812 ◽  
Author(s):  
Martha Sue Carraway ◽  
Andrew J. Ghio ◽  
Jacqueline D. Carter ◽  
Claude A. Piantadosi
Keyword(s):  
Enzyme Activity ◽  
Western Blot ◽  
Heme Oxygenase ◽  
Chronic Hypoxia ◽  
Weight Ratio ◽  
Blood Gases ◽  
Heart Weight ◽  
Heme Oxygenase 1 ◽  
Sprague Dawley ◽  
Control Value

Heme oxygenase (HO)-1 is an oxygen-dependent enzyme that may regulate vascular tone and cell proliferation through the production of carbon monoxide (CO). We tested the hypothesis that HO-1 is upregulated in the lung in chronic hypoxia by exposing male Sprague-Dawley rats to 17,000 feet (395 Torr) for 0, 1, 3, 7, 14, or 21 days. After exposure, blood gases, carboxyhemoglobin (COHb) levels, and hematocrit were measured, and the lungs were either inflation fixed for immunohistochemistry or frozen for later measurement of HO enzyme activity, Western blot for HO-1 protein, and RT-PCR for HO-1 mRNA. The heart was excised and weighed, and the right-to-left heart weight ratio was determined. During hypoxia, the hematocrit increased progressively, reaching significantly higher values than the control value after 3 days. COHb levels increased above the control value after 1 day of hypoxia and increased progressively between 14 and 21 days, whereas arterial[Formula: see text] and arterial[Formula: see text] did not vary significantly. HO-1 protein determined by Western blot increased for the first 7 days and declined thereafter; however, enzyme activity was elevated only after 1 day. Changes in HO-1 during hypoxia were localized by immunohistochemistry to inflammatory cells (early) and newly muscularized arterioles (later). Lung HO-1 mRNA normalized to glyceraldehyde-3-phosphate dehydrogenase was increased after 1 and 21 days. The data indicate that lung HO-1 protein and activity are upregulated only during early chronic hypoxia, whereas persistent COHb elevations indicate high endogenous CO production rates at nonpulmonary sites. If CO has antiproliferative properties, the lack of HO enzyme activity in the lung may be permissive for pulmonary vascular proliferation in hypoxia.

scholarly journals Nutritional and hormonal regulation of malic enzyme synthesis in rat mammary gland

1986 ◽  
Vol 236 (2) ◽  
pp. 441-445 ◽  
Author(s):  
M F Lobato ◽  
M Ros ◽  
F J Moreno ◽  
J P García-Ruíz
Keyword(s):  
Enzyme Activity ◽  
Mammary Gland ◽  
Malic Enzyme ◽  
Hormonal Regulation ◽  
Relative Rate ◽  
Enzyme Synthesis ◽  
Soluble Proteins ◽  
Control Value ◽  
Rat Mammary Gland ◽  
Malic Enzyme Activity

Cytosolic malic enzyme was purified from rat mammary gland by L-malate affinity chromatography. The pure enzyme obtained was used to produce a specific antiserum in a rabbit. Relative synthesis of malic enzyme in the mammary gland of mid-lactating rats was 0.097%, measured by labelling the enzyme in isolated acini. When food was removed, malic enzyme synthesis decreased to 35% and 20% of the control value at 4 and 6 h respectively. Incorporation of [3H]leucine into soluble proteins was constant during the first 6 h of starvation. When lactating rats (maintained with their pups) were starved for 24 h and then re-fed, the relative rate of enzyme synthesis increased 2.5-, 4-, and 4.5-fold at 3 h, 6 h and 18 h respectively after initiation of re-feeding. The relative rate of malic enzyme synthesis was about 50% of normal at 15 h after weaning, whereas the rate of synthesis of soluble proteins did not change. Administration of bromocriptine or adrenalectomy of lactating rats decreased the relative rate of synthesis of malic enzyme by 40% or 30% respectively; these effects were counteracted by hormone supplementation. Hormone therapy also caused an increase in the rate of incorporation of [3H]leucine into soluble proteins and in malic enzyme activity.

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