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1.
L M Gubernieva E E Safronova G A Annenkov V N Malakhov V B Mammaev 《Biokhimii?a (Moscow, Russia)》1976,41(1):153-160
A kinetic method of estimating the ratio of mole quota of H and M human lactate dehydrogenase (LDG) subunits is proposed, based on changes in substrate inhibition of LDG isoenzymes with lactate. Stability of kinetic constants for a long period of time is demonstrated. The dependency of activities ratio under low and high substrate concentration on the contribution of mole quota of LDG M subunits is studied. The correlation of experimental and theoretical values is shown to be: r=0.998 p less than 0.001. A comparison is carried out of the content of LDG subunits molar quotas in artificial mixtures with electrophoretic experimental data, a good coinsidence of these values being registered. The informative importance of the method described with standard methods of the estimation of LDG isoenzyme systems is discussed. No effect of components of human diploid cells homogenate and an insignificant effect of blood serum components on kinetic constants of LDG isoenzymes is registered. A dependency of variation coefficients on the enzyme activity is studied, minimal omegan value being 0.6%. The applicability of the method described for the calculation of quantitative content of both LDG subunits in natural objects (blood serum, diploid cell homogenate etc.) is demonstrated. 相似文献
2.
L. M. Gubernieva E. E. Safronova V. N. Malakhov V. B. Mamaev G. A. Annenkov 《Molecular and cellular biochemistry》1978,19(3):165-170
Summary A kinetic method of estimating the mole quota ratios of the human lactate dehydrogenase (LDH) H and M subunits based on differences in substrate inhibition of LDH isoenzymes by lactate is proposed. Stability of kinetic constants for a prolonged period of time is demonstrated. The dependence of the activity ratios on the contribution of the mole quota of the M-subunit of LDH is studied under conditions of low and high substrate concentrations. The experimental and theoretical values show the following correlation: r = 0.998; p < 0.001. A comparison of the method proposed with the electrophoretic method of LDH subunit estimation is made, the values obtained being in good agreement. No effect of the components of human diploid cell homogenate and only an insignificant effect of the blood serum components on the kinetic constants of LDH isoenzymes are shown. The applicability of the method to the estimation of the quantitative content of both LDH subunits in natural samples is demonstrated. The informational value of the method is compared to that of other standard methods of LDH isoenzyme estimation.The need of the rapid and reliable method for determination the lactate dehydrogenase (LDH) activity of the H and M subunits has long been a matter of great importance, since the study of LDH isoenzymes is an indispensable part of clinical, genetical, cytological and herontological investigations.In 1960 PLAGEMANN et al.
1
,making use of different substrate inhibition of H4 and M4 isoenzymes LDH, developed a method for the estimation of the percent composition H and M subunits LDH within any given mixture of them. The method involves the assay of mixture of LDH isoenzymes in the presence of two different levels of pyruvate. The authors calculated the percent of each subunit in a mixture from the ratio of enzymatic activities at both high and a low concentration of pyruvate. Although this method was subsequently improved, both experimentally2–S and theoretically6, its application was still impossible without first eliminating a great many problems. The problem of subunit interactions inside the enzyme molecule has not been settled. In addition, questions have not been raised about stability of the kinetic parameters', the reproducibility of the method, its applicability to the study of different objects and also the informational value of the experimental data.In our previous investigation7,8, we have studied the kinetic properties of five purified isoenzymes of human lactate dehydrogenase and demonstrated the catalytic independence of the active sites of the LDH tetrameric molecules with respect to substrate inhibition.In the present report an attempt has been made to develop a kinetic method for the assay of M-polypeptide chains mole quotum of lactate dehydrogenase in natural specimens. 相似文献
3.
The activity of cytoplasmic superoxydase (SOD-1) was studied in erthrocytes of 17 patients affected with Down's syndrome (trisomy 21) and in 26 healthy persons. A 1.56-fold increase of the enzyme activity was observed in the group of patients as compared with the control group. This could be explained as the dosage effect of the corresponding gene located in the chromosome 21. 相似文献
4.
D N Propert 《Human heredity》1979,29(6):361-363
Pseudocholinesterase activity and the phenotypes controlled by the E1 locus have been determined in a sample of 307 Down's syndrome patients and 206 patients suffering from nonspecific mental retardation and compared to those in the healthy population. Both groups of patients have an elevated frequency of phenotypes possessing the rate E1f allele. The mentally retarded patients have a higher mean pseudocholinesterase activity than those with Down's syndrome who, in turn, have activity than the healthy controls. 相似文献
5.
Average lactate dehydrogenase (LDH) isoenzyme patterns the content of H subunits, total LDH activity, total malate dehydrogenase (MDH) activity and the m- MDH/s-MDH ratio were determined in twelve muscles and the male genital tract of the rabbit. LDH(1) was the predominant form in the heart, soleus and masseter muscles, LDH(3) in the lingual muscles and LDH(5) in the other muscles analysed. In the muscles, an increase in the percentual proportion of M subunits was accompanied, by a proportional increase in total LDH activity and a decrease in total MDH activity, especially m-MDH. LDH isoenzyme patterns and LDH and MDH activities are useful for obtaining some idea about the proportion of individual muscle fibres. Activity accounted for by H subunits was roughly the same in all the muscles analysed, indicating that the synthesis of H subunits is independent of the type of muscle fibre and of the oxygen supply of the muscular tissue, and also that isoenzymes composed chiefly of H subunits are not localized preferentially in the mitochondria. Similar relationships between LDH isoenzymes and LDH and MDH activities were found in the testicular and epididymal tissues. The tests and the head of the epididymis mainly contain LDH isoenzymes composed of H subunits. The total LDH activity in these tissues is relatively low and their MDH activity is relatively high compared with the body and tail of the epididymis. The proportion of H subunits in the ampulla, the seminal vesicles, the coagulating glands and the prostate is also high. Cowper's glands have a high LDH(5) and LDH(4) concentration. One of two LDHx isoenzymes were found in the testes and spermatozoa. 相似文献
6.
Summary A number of enzymes alkaline and acid phosphatase, lactic acid dehydrogenase, glutamic oxalacetate transaminase (GOT), aldolase, glucose-6-phosphat dehydrogenase (G-6-PD) in the serum of 81 patients with Langdon Down's syndrome and mental deficiency (idiocy) were investigated and then compared with a sample of 37 healthy persons. The enzyme activities were determined spectrophotometrically.Statistically significant differences of alkaline and acid phosphatase, lactic acid dehydrogenase, and glutamic oxalacetate transaminase were demonstrated between oligophrenic patients and healthy persons; there was a distinct increase in the serum of patients. The values for aldolase and glucose-6-phosphat dehydrogenase were equal both in the serum of patients and of controls. There was no significant difference between the values of adult oligophrenics on one side and adult patients with Down's syndrome on the other. Until further notice children with serious mental deficiency — oligophrenics of unknown genesis as well as patients with Down's syndrome — demonstrated no definite evidence of enzymologically deviations compared with those of the control group.
(Direktor: Prof. Dr. med. H. Schade)
Nach einem am 22. 10. 1967 auf der 10. Tagung der Gesellschaft für Anthropologie und Humangenetik gehaltenen Vortrag. 相似文献
(Direktor: Prof. Dr. med. H. Schade)
Nach einem am 22. 10. 1967 auf der 10. Tagung der Gesellschaft für Anthropologie und Humangenetik gehaltenen Vortrag. 相似文献
7.
Red blood cell glucose metabolism in Down's syndrome 总被引:1,自引:0,他引:1
M Magnani V Stocchi G Novelli M Dachà G Fornaini 《Clinical physiology and biochemistry》1987,5(1):9-14
The specific activity of red blood cell glycolytic enzymes was determined in 20 Down's syndrome patients and compared with 20 normal controls. According to previous evidence, a 50% increase of phosphofructokinase and a 30% increase of glucose-6-phosphate dehydrogenase and glutathione peroxidase activity was found. Metabolic studies of the patients' erythrocytes revealed a decrease in fructose-6-phosphate and 2, 3-diphosphoglycerate concentrations, while fructose-1, 6-diphosphate and ADP both increased. Glucose utilization by intact erythrocytes from Down's syndrome patients did not differ from that of normal controls. However, addition of methylene blue or inorganic phosphate produced a higher stimulation of erythrocyte glycolysis in patients with Down's syndrome compared to controls. These metabolic abnormalities could be, at least in part, ascribed to the increased phosphofructokinase activity which is due to a gene-dosage effect. 相似文献
8.
T Miyakawa K Watanabe S Katsuragi 《Virchows Archiv. B, Cell pathology including molecular pathology》1986,52(2):99-106
Amyloid fibrils in brains of patients with Alzheimer's disease and Down's syndrome were examined by light and electron microscopy. In addition, replicas of amyloid fibrils produced by a quick freezing method from the brain of a patient with Down's syndrome were examined by electron microscopy. The amyloid fibrils were shown to consist of hollow rods. These were composed of filaments arranged as a tightly coiled helix, each turn of which consisted of five globular subunits. This structure appears to be similar to the prion filament observed in Creutzfeldt-Jakob disease (CJD). The possibility therefore arises that amyloid fibrils in Alzheimer's disease and Down's syndrome may be related to the transmissible agents responsible for diseases such as CJD, kuru and Gerstmann-Str?ussler Syndrome (GSS). 相似文献
9.
Structural and functional characterization of the inhibition of urokinase by alpha 2-macroglobulin 总被引:3,自引:0,他引:3
We have investigated the interaction of alpha 2-macroglobulin (alpha 2M) with the serine proteinase urokinase, an activator of plasminogen. Urokinase formed sodium dodecyl sulfate stable complexes with purified alpha 2M and with alpha 2M in plasma. These complexes could be visualized after polyacrylamide gel electrophoresis by protein blots using 125I-labeled anti-urokinase antibody or by fibrin autography, a measure of fibrinolytic activity. According to gel electrophoretic analyses under reducing conditions, urokinase cleaved alpha 2M subunits and formed apparently covalent complexes with alpha 2M. Urokinase cleaved only about 60% of the alpha 2M subunits maximally at a mole ratio of 2:1 (urokinase: alpha 2M). Binding of urokinase to alpha 2M protected the urokinase active site from inhibition by antithrombin III-heparin and inhibited, to a significant extent, plasminogen activation by urokinase. Reaction of urokinase with alpha 2M caused an increase in intrinsic protein fluorescence and, thus, induced the conformational change in alpha 2M that is characteristic of its interactions with active proteinases. Our results indicate that both in plasma and in a purified system the alpha 2M-urokinase reaction is functionally significant. 相似文献
10.
Differential turnover of phosphate groups on neurofilament subunits in mammalian neurons in vivo 总被引:11,自引:0,他引:11
The phosphorylation and dephosphorylation of specific proteins was demonstrated directly in the intact vertebrate nervous system in vivo. By exploiting the neurons' ability to segregate a select group of cytoskeletal proteins from most other phosphorylated constituents of the cell by axoplasmic transport, we were able to examine the dynamics of phosphate turnover on neurofilament proteins in mouse retinal ganglion cell neurons simultaneously labeled with [32P]orthophosphate and [3H]proline in vivo. Three [3H]proline-labeled neurofilament protein (NFP) subunits, designated H (160-200 kDa), M (135-145 kDa), and L (68-70 kDa), entered optic axons in a mole:mole ratio similar to that of isolated axonal neurofilaments, supporting the notion that newly synthesized NFPs are transported along axons as assembled neurofilaments. NFP subunits incorporated high levels of 32P before reaching axonal sites at the level of the optic nerve. As neurofilaments were transported along axons, however, many initially incorporated [32P]phosphate groups were removed. Loss of these phosphate groups occurred to a different extent on each subunit. A minimum of 50-60 and 35-40% of the labeled phosphate groups was removed in a 5-day period from the L and M subunits, respectively. By contrast, the H subunit exhibited relatively little or no phosphate turnover during the same period. Dephosphorylation of L in axons is accompanied by a decrease in its net state of phosphorylation; changes in the phosphorylation state of H and M, however, also reflect ongoing addition of phosphates to these polypeptides during axonal transport (Nixon, R.A., Lewis, S.E., and Marotta, C.A. (1986) J. Neurosci., in press). The possibility is raised that dynamic rearrangements of phosphate topography on NFPs represent a mechanism to coordinate interactions of neurofilaments with other proteins as these elements are transported and incorporated into the stationary cytoskeleton along retinal ganglion cell axons. 相似文献
11.
Isoenzyme patterns, total lactate dehydrogenase (LDH, E.C. 1.1.1.27) activity and H and M subunit activity were determined in the tissues of Czech Spotted bovine foetuses. Total LDH activity rose in the skeletal muscles throughout the whole of the prenatal period. In the viscera it usually attained the maximum at a foetal length of 66.7 cm. Differences in the isoenzyme patterns of the various organs of an 8.1-cm foetus were relatively small (41.9--66.1% H subunits). In the heart and kidneys, in which LDH1 and LDH2 markedly predominate in adulthood, the isoenzyme pattern resembled the adult one at a length of only 13.3 cm, but in the liver, spleen and lungs not until 66.7 cm. The proportion of H subunits also rose in the part of the gastrointestinal tract where secretory and resorptive activity predominate (the abomasum, the small and the large intestine). Conversely, it fell in organs concerned mainly with the mechanical processing of food (the rumen, reticulum and omasum). The proportion of M subunits rose in all the skeletal muscles up to a foetal length of 66.7 cm. Later on, differentiation into muscles in which M subunits predominated (the longissimus dorsi and the triceps brachii), into muscles with approximately the same proportion of H and M subunits (the iliopsoas) and to muscles with a preponderance of H subunits (the masseter and the muscular part of the diaphragm) occurred. 相似文献
12.
Trisomy 21 (Down's syndrome) is the most common genetic cause of human mental retardation. In Down's syndrome (DS) patients, deteriorated glucose, lipid, purine, folate and methionine/homocysteine metabolism has been reported. In our study, we used a proteomic approach to evaluate protein expression of enzyme proteins of intermediary metabolism in the brain of Down's syndrome fetuses. In fetal DS brain, we detected increased protein levels of mitochondrial aconitase as well as NADP-linked isocitrate dehydrogenase, decreased protein expression of citrate synthase and cytosolic aspartate aminotransferase. From two spots that corresponded to either pyruvate kinase M1 or M2 isozymes, significant elevation was observed only in one, while the second spot as well as the sum of the spots showed no differences between DS and controls. These results suggest derangement of intermediary metabolism during prenatal development of DS individuals. 相似文献
13.
Four cases of Down's syndrome in new-born have been thoroughly studied on the histological and morphometrical point of view. It is outlined the presence in the seminiferous tubules of abnormal spermatogonia, sometimes multinucleated and characterized by a prominent nucleolus. The Authors emphasize the conspicuous number of these hypertrophic (H) spermatogonia (usually very scanty in normal new-born) never reported in the Down's syndrome before, whose significance is not easy to explain: if they represent anomalous forms of spermatogonial cells, related to the malformative condition, or a premalignant aspect is then discussed. Morphometrical measurements of the average tubular diameter (D.T.M.) and spermatogonia content have been calculated; furthermore a microprocessor system is employed for evaluating nuclear diameter, perimeter and area not only in H spermatogonia, but also in the other germ cells observed (A and P). 相似文献
14.
Down's syndrome: abnormal neuromuscular junction in tongue of transgenic mice with elevated levels of human Cu/Zn-superoxide dismutase 总被引:17,自引:0,他引:17
To investigate the possible involvement of Cu/Zn-superoxide dismutase (CuZnSOD) gene dosage in the neuropathological symptoms of Down's syndrome, we analyzed the tongue muscle of transgenic mice that express elevated levels of human CuZnSOD. The tongue neuromuscular junctions (NMJ) in the transgenic animals exhibited significant pathological changes, namely, withdrawal and destruction of some terminal axons and the development of multiple small terminals. The ratio of terminal axon area to postsynaptic membrane decreased, and secondary folds were often complex and hyperplastic. The morphological changes in the transgenic NMJ were similar to those previously seen in muscles of aging mice and rats as well as in tongue muscle of patients with Down's syndrome. The findings suggest that CuZnSOD gene dosage is involved in the pathological abnormalities of tongue NMJ observed in Down's syndrome patients. 相似文献
15.
A P Akif'ev G R Mutovin I V Filiushkin E K Khandogina 《Biulleten' eksperimental'no? biologii i meditsiny》1983,96(12):69-71
A study was made of the yield of chromosome aberrations in gamma-irradiated G0 peripheral blood lymphocytes from 6 patients with different forms of Down's syndrome. The doses used were from 0.25 to 3.0 Gy. Seven healthy donors of different age made the control group. There was a significant increase in the yield of chromosome exchanges in lymphocytes from all the patients as compared to control. The spontaneous level of chromosome aberrations and the frequency of radiation-induced fragments did not differ from the control values. The yield of exchanges in diploid and trisomic cells from patients with the mosaic form of Down's syndrome did not change significantly as the time of cultivation was raised. The origin of DNA repair defects leading to the increased chromosome radiosensitivity in Down's syndrome is discussed. 相似文献
16.
Migliore L Botto N Scarpato R Petrozzi L Cipriani G Bonuccelli U 《Cytogenetics and cell genetics》1999,87(1-2):41-46
To further investigate our finding of high levels of spontaneous aneuploidy in somatic cells of Alzheimer's disease (AD) patients (Migliore et al. 1997), we studied the molecular cytogenetics of eight patients with sporadic AD and six healthy controls of similar age. Cytochalasin B-blocked binucleated peripheral blood lymphocytes from the AD patients and unaffected controls were used to measure micronucleus induction or other aneuploidy events, such as the presence of malsegregation in interphase nuclei (representing chromosome loss and gain). Dual-color fluorescence in situ hybridization (FISH) with differential labeled DNA probes was applied. We used a probe specific for the centromeres of chromosomes 13 and 21 combined with a single cosmid for the Down's syndrome region (21q22.2) to obtain information on spontaneous chromosome loss and gain frequencies for both chromosomes (13 and 21). FISH data showed that AD lymphocytes had higher frequencies of chromosome loss (evaluated as fluorescently labeled micronuclei) for both chromosomes, as well as higher frequencies of aneuploid interphase nuclei, again involving both chromosomes, compared to control lymphocytes. However, aneuploidy for chromosome 21 was more frequent than for chromosome 13 in AD patients. This preferential occurrence of chromosome 21 in malsegregation in somatic cells of AD patients raises the hypothesis that mosaicism for trisomy of chromosome 21 could underlie the dementia phenotype in AD patients, as well as in elderly Down's syndrome patients. 相似文献
17.
In the horse, total LDH activity increased with training and the H and M subunit activity parallelled this increase. It is suggested that these increases are in response to a stimulus from the type of training program utilised. The first half of a detraining program decreased the activity of the H and M subunits as might be expected. A sharp rise in the total LDH and the M subunit activity occurred during the latter half of the detraining program. This unexpected increase may be due to relatively more hypoxic conditions prevailing in the muscle during the detraining period. 相似文献
18.
Novobiocin precipitates histones at concentrations normally used to inhibit eukaryotic type II topoisomerase. 总被引:6,自引:7,他引:6
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At concentrations normally used to inhibit eukaryotic type II topoisomerase activity (100-1000 micrograms/ml) novobiocin binds core histones. Approximately 15 moles of novobiocin bind per mole of histone resulting in histone precipitation from solution in either 0.15 M or 2 M NaCl. The interaction between novobiocin and proteins appears to involve arginine residues: histones H3 and H4 (13.5 and 14 mole percent arginine) are precipitated at lower novobiocin concentrations than histones H2A and H2B (9.5 and 6.5 mole percent arginine). Furthermore, polyarginine but not polyornithine competes for novobiocin in histone precipitation. Moreover, histones with arginine residues modified with 1,2-cyclohexanedione are soluble in 1000 micrograms/ml novobiocin. Because novobiocin can remove histones from solution as well as inhibit topoisomerase activity, and because both of these events can alter DNA topology, novobiocin should be used with caution in experiments designed to implicate topoisomerase activity in chromatin dynamics. 相似文献
19.
20.
Increasing evidence of the interaction of glucocorticoids and ovarian steroids prompted the current study. Effects of exogenously administered corticosterone acetate (3.5 mg/100 g b.w/day for one week) were examined on splenic nucleic acids, protein, lactate, and on lactate dehydrogenase (LDH) specific activity and its isozymes in ovariectomized and ovary-intact Wistar rats (65-75 days old). Ovariectomy resulted in no significant change in the parameters studied except DNA which increased significantly. The administration of corticosterone to these rats did not produce any remarkable change in the ovariectomy caused increase in splenic DNA content. Nevertheless, it decreased the ratio of heart type subunits (H)/muscle type subunits (M) [H/M] of LDH isozymes. In the case of ovary-intact rats, corticosterone produced an increase in the concentration of splenic lactate but a decrease in the H/M ratio. Exogenously administered corticosterone exerts selective synergistic interaction with ovarian hormones on splenic lactate. The specific activity of LDH and the concentrations of RNA and protein remained unchanged during the interaction between ovarian hormones and corticosterone. 相似文献