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PubMed1998

[Genetic aspects of silicosis: polymorphic gene distribution frequency].

Electrophoresis and isoelectrofocusing were used to study polymorphism by 7 genetic loci: haptoglobin (Hp), proteinase inhibitor (PI), transferrin (TF), Vitamin D-transporting protein (GC), complement 3 (C3), phosphoglucomutase 1 (PGM1) and glyoxalase (GLO1) in 60 patients with silicosis and in 70 apparently healthy workers of the Dynamo plant. Comparison of the study groups by significant differences in the summary of the genetic information obtained suggests that 5 (Hp, C3, TF, PI, PGM1) of the 7 studied systems showed the hereditary features of silicosis. The gene carriers Hp*2, C3*F, PGM1*2-, PI*M1, TF*C1, TF*C16 TF*D, GC*R due to peculiar biochemical processes appear to have less adaptive potentialities and a greater likelihood of the disease on exposure to industrial factors.

Adult↗

PubMedDate not supplied

[Enzymatic profile of the exudate leukocytes in diabetes mellitus].

The activity of enzymes regulating the processes providing functional activity of leukocytes was studied in the exudate leukocytes of healthy rabbits and animals with alloxan diabetes. Rabbits with diabetes displayed a reduction of hexokinase, phosphoglucomutase, glucose-6-phosphate dehydrogenase and adenylate kinase activity. The activity of UDPH-pyrophosphorylase, UDPH-glycogentranspherase, 6-phosphogluconate dehydrogenase and glutathion reductase showed no significant changes in the exudate leukocytes in diabetes. A reduction of hexokinase and glucose-6-phosphate dehydrogenase limiting glycolysis and the pentose-phosphate cycle, respectively, providing energy for leukocytes and important in protein metabolism of these cells, is of great significance in the reduction of functional activity of leukocytes in the inflammatory focus in diabetes.

Adenylate Kinase↗

PubMed1989

The phosphoglycerate mutases.

The phosphoglycerate mutase family is generally very well documented with respect to structure, evolution, and mode of action. However, a few individuals in the family remain relatively poorly characterized and will clearly require more detailed study. Furthermore, certain aspects of the detailed behavior of these enzymes are, as yet, incompletely understood and require further investigation. Cofactor-dependent monophosphoglycerate mutase and bisphosphoglycerate mutase are undoubtedly very closely related. Their amino acid sequences are strongly similar, they can form active heterodimers, and they catalyze the same three reactions, albeit at substantially different relative rates. Both enzymes catalyze a ping-pong type of reaction with a phosphohistidine intermediate. The presence of an additional phospho ligand at the active site of monophosphoglycerate mutase helps to explain why this enzyme is better at retaining the 2,3-bisphosphoglycerate intermediate and why it is thus more efficient (by a factor of about 10(3)) at catalyzing the interconversion of 3- and 2-phosphoglycerates. The reason why 1,3-bisphosphoglycerate is a better substrate for bisphosphoglycerate mutase than for monophosphoglycerate mutase (by a factor of about 30) is not yet apparent but presumably relates to the relative positioning of the two phospho-binding sites. Both enzymes are equally good as phosphatases when the reaction is activated by 2-phosphoglycollate. Available evidence indicates that these mutases are similar in many respects to the much smaller, cofactor-dependent monophosphoglycerate mutase from Schizosaccharomyces pombe, but further information is required to define the relationship more precisely. Cofactor-independent monophosphoglycerate mutase belongs to a quite distinct branch of the phosphoglycerate mutase family. It is not known at present whether this branch is related divergently or convergently to the cofactor-dependent monophosphoglycerate mutase/bisphosphoglycerate mutase branch. Existing evidence can be argued both ways. For example, the kinetic evidence shows a ping-pong type of reaction and would be consistent with a phosphohistidine intermediate as encountered in the other mutases. Thus the cofactor-independent enzyme may also have arisen by gene duplication--but, in this case, yielding an enzyme of about twice the size, with slightly different residues at the active site and C-terminal tail. An alternative possibility, of course, is that the two branches of the phosphoglycerate mutase family are quite unrelated in a divergent sense and are little more similar structurally than is, for example, the catalytically similar enzyme phosphoglucomutase.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1995

Myoepithelial cell differentiation in the developing mammary gland: progressive acquisition of smooth muscle phenotype.

The most important portion of the mammary gland development occurs postnatally, with distinct periods of intensive morphogenesis taking place between birth and puberty and during pregnancy and lactation. To characterize the differentiation process of mammary myoepithelial cells, we have studied the expression patterns of several smooth muscle phenotypic markers, including contractile proteins, alpha-smooth muscle-actin (alpha-SM-actin), smooth muscle myosin heavy chains (SM-MHC), and calponin; components of cell-extracellular matrix adherens junctions, phosphoglucomutase-related protein (PGM), vinculin variants, integrin subunits; and laminin variant chains in the developing rat mammary gland using immunofluorescence microscopy. alpha-SM-actin- and SM-MHC-positive cells were found first in newborn animals, while calponin, PGM and alpha1 integrin subunit began to be expressed in prepubertal animals (1.5 weeks). Vinculin, beta1 and alpha3 integrin subunits were largely confined to the basal cell layer at all developmental stages examined with greater staining starting at 1.5 weeks. Meta-vinculin was identified only in myoepithelial cells of the lactating gland. gamma1 laminin chain was present in the mammary gland basement membrane throughout development, while the beta2 chain was revealed in 3-week-old animals and accumulated later in postpubertal animals (7 weeks). Similarly, beta2 laminin chain was absent from the forming alveoli basement membrane in pregnant rats and started to accumulate in the lactating gland. In addition to temporal changes, we have observed spatial differences in the distribution of the phenotypic markers. Both in pre- and in postpubertal animals, alpha-SM-actin- and SM-MHC-positive cells of the growing ductal ends contained low amounts if any of calponin, PGM, and beta2 laminin chain. We conclude that during postnatal development, mammary myoepithelial cells progressively acquire a differentiated phenotype as revealed by the expression of various smooth muscle markers. Maturation of the myoepithelial cells is accompanied by upregulation of the smooth muscle integrin expression followed by accumulation of beta2-containing laminin variant. Thus, changes in adhesion system parallel with the myoepithelial cell differentiation.

Animals↗

PubMed1980

Genetic polymorphisms in isolated Sherpa populations of Nepal.

Data are reported on the incidence of variants in the ABO blood group system, four red cell enzyme polymorphisms, and the PTU taster/non-taster polymorphism in 155 Sherpas and Tamangs living in two villages in the isolated Rolwaling valley in east Nepal. The incidence of phenotype AB was unexpectedly high in one village, as has been reported in some other high-altitude populations: possible causes are discussed. Each of the other polymorphisms has a different pattern of distribution between the populations studied; the Sherpas' allele distribution in the phosphoglucomutase (PGM) system seems to be distinct from the surrounding populations'. ABO gene frequencies suggest affinities between Sherpas in the study villages and old- and new-clan Sherpa populations, respectively, in Solu and Khumbu.

ABO Blood-Group System↗

PubMed1981

Reindeer Chukchi and Siberian Eskimos: studies on blood groups, serum proteins, and red cell enzymes with regard to genetic heterogeneity.

Phenotype and allelic frequencies for ABO, MNSs, P, Rh, Kell, Duffy, and Diego blood groups, as well as for transferrins (Tf), haptoglobins (Hp), phosphoglucomutase 1 (PGM1), adenylate kinase (AK), 6-phosphogluconate dehydrogenase (PGD), and acid phosphatase (AcP) are described in 9-10 adjoining populations of Reindeer Chukchi. Additionally, one of three presently existing territorial subgroups of Siberian Eskimos was studied. The total sample size ranges from 931 to 1,066 in Chukchi, and from 99 to 102 in Eskimos, depending on the genetic system studied. Substantially reduced samples for Kidd and ABO secretion were investigated solely in the Reindeer Chukchi. Significant heterogeneity of allelic frequencies has been observed among Chukchi populations (X(2)124 = 378.47, P less than 0.001). Summed genetic heterogeneity between Reindeer Chukchi and Siberian Eskimos was also found to be highly significant (X(2)14 = 186.54, P less than 0.001). Both groups can be readily discriminated with only four outliers: Ns, P1, R1, and pa. Random genetic drift is suggested to be responsible for a large proportion of heterogeneity of allelic frequencies at the MNSs, P, and Rh blood-group system sites both among the Chukchi and Eskimos. Conversely, stabilizing selection is assumed as the principal agent maintaining homogeneous allelic frequencies at the AcP locus within the Chukchi subdivisions, whereas disruptive selection may be considered as a major factor leading to different Pa frequencies between Chukchi and Eskimos.

Arctic Regions↗

PubMed1983

Genetic factors in the population of Plati, Greece.

One-thousand, thirty-eight individuals from Plati, Greece were examined for the following red cell antigens, serum proteins, and red cell enzymes A A1 Ai B H; MNSs Mg Henshaw Nya Mur Vw; CCwcDEeCe; K k Kpa Kpb Jsa Jsb; P1; Lua; Fy1 Fy2; Jka Jkb; Wra; Zt; Vel; Swa; Jensen, Radin, Gerbich, Diego, Gregory, Haptoglobin, Transferrin, Acid phosphatase, Adenylate kinase, Adenosine deaminase, Esterase-D, Glucose-6-phosphate dehydrogenase, Phosphoglucomutase, 6-Phosphogluconate dehydrogenase, Phosphohexose isomerase, Lactate dehydrogenase, Malate dehydrogenase, and Superoxide dismutase. The results are discussed in detail and compared with other Greek and neighbouring populations. Because of the Plati population's long history of residence in the Cappadocian area of Turkey the data have been compared, whenever possible, with results for that region.

Adolescent↗

PubMed1988

Blood genetic markers in Sri Lankan populations--reappraisal of the legend of Prince Vijaya.

Serum protein (haptoglobin types; transferrin and group-specific component subtypes); haemoglobin and red cell enzymes (acid phosphatase, esterase D, glyoxalase I, 6-phosphogluconate dehydrogenase, adenylate kinase, and phosphoglucomutase (locus 1) (subtypes) were studied in the Sinhalese, Tamils, and Muslims of Sri Lanka. The allelic frequencies of all the polymorphic systems were similar in these populations without any significant differences. A close look at the present results and earlier investigations on 13 polymorphic loci controlled by 37 alleles did not reveal any genetic characteristics in the present-day Sinhalese population that are distinct from those in the Tamils of Sri Lanka. As such, genetic evidence linking the legendary origin of the Sinhalese population to East India (Prince Vijaya) is lacking.

Blood Proteins↗

PubMed2005

Dynamics of intracellular metabolites of glycolysis and TCA cycle during cell-cycle-related oscillation in Saccharomyces cerevisiae.

In the present work LC-MS/MS was applied to measure the concentrations of intermediates of glycolysis and TCA cycle during autonomous, cell-cycle synchronized oscillations in aerobic, glucose-limited chemostat cultures of Saccharomyces cerevisiae. This study complements previously reported oscillations in carbon dioxide production rate, intracellular concentrations of trehalose and various free amino acids, and extracellular acetate and pyruvate in the same culture. Of the glycolytic intermediates, fructose 1,6-bisphosphate, 2- and 3-phosphoglycerate, and phosphoenolpyruvate show the most pronounced oscillatory behavior, the latter three compounds oscillating out of phase with the former. This agrees with previously observed metabolic control by phosphofructokinase and pyruvate kinase. Although individually not clearly oscillating, several intermediates of the TCA cycle, i.e., alpha-ketoglutarate, succinate, fumarate, and malate, exhibited increasing concentration during the cell cycle phase with high carbon flux through glycolysis and TCA cycle. The average mass action ratios of beta-phosphoglucomutase and fumarase agreed well with previously determined in vitro equilibrium constants. Minor differences resulted for phosphoglucose isomerase and enolase. Together with the observed close correlation of the pool sizes of the involved metabolites, this might indicate that, in vivo, these reactions are operating close to equilibrium, whereby care must be taken due to possible differences between in vivo and in vitro conditions. Combining the data with previously determined intracellular amino acid levels from the same culture, a few clear correlations between catabolism and anabolism could be identified: phosphoglycerate/serine and alpha-ketoglutarate/lysine exhibited correlated oscillatory behavior, albeit with different phase shifts. Oscillations in intracellular amino acids might therefore be, at least partly, following oscillations of their anabolic precursors.

Cell Cycle↗

PubMed1988

Isoenzyme analysis of lichen algae in immobilized pH gradients.

A base for a modern species' concept of chlorococcal algae can be obtained not by morphological analysis, but by biochemical characters, e.g. isoenzyme banding patterns. From isolated lichen algae of the genus Trebouxia de Puymaly a set of five such enzymes has been studied by isoelectric focusing in immobilized pH gradients (IPG): phosphoglucomutase, phosphoglucose isomerase, malate dehydrogenase, mannitol dehydrogenase and leucine aminopeptidase. The first four are resolved into isoforms in a pH 4-7 IPG interval, while the last one is analyzed in an IPG pH 3.5-5 span. The patterns are specific for distinct populations, inter- and intraspecifically varying in dependence from their geographical distribution or the lichen species from which they have been isolated. Their limited heterogeneity (one to four isoforms) suggests that they are the products of specific genes rather than artefacts of the extraction procedure or the IPG analysis. Sharp isozyme patterns can only be obtained in a mixed-bed, carrier-ampholyte (CA)-IPG gel and by anodic application, suggesting that the recently proposed mechanism of hydrophobic protein-IPG matrix interaction (Electrophoresis, 1987, 8, 62-70) is fully operative here. As an additional mechanism, it is proposed that, in some cases, CA might simply act, when added to an IPG gel, by buffering, in the transient state, the sample zone before the protein migrates from the liquid phase into the IPG matrix.

Ampholyte Mixtures↗

PubMed1992

The use of immobilized pH gradients for the detection of human polymorphisms in the forensic identification of bloodstains.

Some polymorphic proteins (alpha 1-antitrypsin, orosomucoid, transferrin, group specific component, plasminogen) and enzymes (phosphoglucomutase, acid phosphatase, estrase D) were determined in bloodstain extracts by isoelectric focusing with carrier ampholytes (CA) and with immobilized pH gradients (IPGs) rehydrated with CA. IPGs yield superior results for typing of genetics markers in bloodstains since phenotypes are better distinguished and the bands are straighter and sharper in the presence of contaminants. Also, the sensitivity of IPGs with CA is similar to isoelectric focusing (IEF) with CA. A new variant, ACP*B1, found in Negroid west African populations and not found in Caucasians is described. Such a variant can only be determined by IPGs since its isoelectric point (pI 5.95) is close to that of the ACP*B (pI 6.05) variant.

Ampholyte Mixtures↗

PubMed1987

Changes in cerebral glycogenolysis and related enzymes in diazinon treated hyperglycaemic animals.

Effect of diazinon (10,20 and 40 mg/kg, i.p.) on the level of blood glucose in rats was investigated. Hyperglycaemia peaked 2 h after i.p. treatment with 40 mg/kg diazinon. The cerebral acetylcholinesterase activity was significantly reduced. The blood level of pyruvic acid was unchanged while that of lactic acid was significantly increased. Convulsions and biochemical changes caused by diazinon (40 mg/kg) were prevented by diazepam injected immediately after diazinon. In diazinon-treated hyperglycaemic animals, the glycogen content of the brain was depleted, the activities of glycogen phosphorylase, phosphoglucomutase and hexokinase were significantly increased and the activity of glucose-6-phosphatase remained unchanged. Lactate dehydrogenase activity was also increased by treatment with diazinon. The induced changes may compensate for the energy requirement of stimulatory effects caused by diazinon.

Animals↗

PubMedDate not supplied

Cerebral glucose and glycogen metabolism in diazinon-treated animals.

The intraperitoneal (IP) treatment of rats with diazinon (40 mg/kg) resulted in a variety of changes in the brain. Glycogen was depleted, but there was an increase in the activities of glycogen phosphorylase, phosphoglucomutase, hexokinase, lactate dehydrogenase, and fructose 1,6 diphosphatase. The activity of glucose-6-phosphatase was unaffected while that of cholinesterase was significantly reduced. Lactic acid content was increased, while that of pyruvate was not altered. Animals developed tremors and convulsions, which were maximal two hours after treatment. The induced changes may be compensatory mechanisms to provide extra energy to cerebral tissue as a result of the stimulatory effects in diazinon-treated animals.

Animals↗

PubMed1979

Enzyme changes accompanying thermal acclimation in two species of pleurocerid snails.

Goniobasis cahawbensis is a stream snail that experiences an annual temperature cycle. G. cochliaris is limited in distribution to springs, and their immediate vicinities, which are characterized by nearly constant annual temperatures. The present study sought to determine whether temperature dependent biochemical differences exist that might account for the differential distribution of these congeneric pleurocerid snails. Eight enzymes were examined following acclimation to 10 degrees, 17 degrees and 24 degrees C. No significant temperature dependent qualitative differences in enzyme phenotypes were demonstrable in either species by starch-gel electrophoresis for malate dehydrogenase, glucose-6-phosphate dehydrogenase, phosphogluconate dehydrogenase, phosphoglucomutase, superoxide dismutase and acetyl and butyryl esterases. Significant quantitative differences were observed in three of these enzymes. G. cahawbensis cytosol malate dehydrogenase activity increased significantly with increasing acclimation temperature, while G. cochliaris malate dehydrogenase activity remained unchanged. The activities of glucose-6-phosphate dehydrogenase did not differ significantly between acclimation temperatures for either species; however, the overall activity of both enzymes was significantly higher for G. cochliaris. Appreciable levels of LDH activity were not demonstrable by electrophoresis or enzymatic assay.

Adaptation, Physiological↗

PubMed2007

Banana (Musa spp.) as a model to study the meristem proteome: acclimation to osmotic stress.

Banana (Musa spp.) multiple shoot meristems are an excellent model to study the meristem proteome. Using a 2-DE protocol developed for small amounts of tissue and MS-based cross species polypeptide identification, we have revealed the meristem proteome and investigated the influence of sucrose-mediated osmotic stress in a dehydration-tolerant variety. Proteins that were significantly up- or down-regulated due to the high-sucrose treatment were classified using non-parametric univariate statistics. Our results suggest that the maintenance of an osmoprotective intracellular sucrose concentration, the enhanced expression of particular genes of the energy-conserving glycolysis and the conservation of the cell wall integrity are essential to maintain homeostasis, to acclimate and to survive dehydration. By comparing the dehydration-tolerant variety with a dehydration-sensitive variety, we were able to distinguish several genotype-specific proteins (isoforms), and could associate the dehydration-tolerant variety with proteins involved in energy metabolism (e.g., phosphoglycerate kinase, phosphoglucomutase, UDP-glucose pyrophosphorylase) and proteins that are associated with stress adaptation (e.g., OSR40-like protein, abscisic stress ripening protein-like protein). This work shows that proteome analysis can be used successfully to perform quantitative difference analysis and to characterize genetic variations in a recalcitrant crop.

Electrophoresis, Gel, Two-Dimensional↗

PubMed2003

Stable isotope labeling of phosphopeptides for multiparallel kinase target analysis and identification of phosphorylation sites.

An approach for multiparallel target identification and relative quantification of in vitro kinase activities in two different biological samples, using liquid chromatography/mass spectrometry (LC/MS), is described. Synthetic target peptides, containing the putative regulatory phosphorylation sites of sucrose-phosphate synthase (SPS) isoenzymes from Arabidopsis thaliana, were simultaneously in vitro phosphorylated and their phosphorylation states determined. Quantification was achieved by stable isotope labeling of the phosphoserine moiety with ethanethiol and [(2)D(5)]-ethanethiol. This revealed different kinase activities in extracts of wild-type (WT) plants and mutant plants lacking plastidic phosphoglucomutase (PGM). The multiparallel assay allowed the determination of favored substrate specificities among the putative phosphorylation sites in SPS. Additionally, we extended the method to unambiguously identify phosphorylation sites in peptides via differential labeling.

Amino Acid Sequence↗

PubMed2004

The trehalose pathway and intracellular glucose phosphates as modulators of potassium transport and general cation homeostasis in yeast.

Trk, encoded by the partially redundant genes TRK1 and TRK2, is the major potassium transporter of Saccharomyces cerevisiae. This system is specific for potassium and rubidium but, by reducing the electrical membrane potential of the plasma membrane, Trk decreases the uptake of toxic cations such as lithium, calcium, aminoglycosides and polyamines, which are transported by other systems. Gain- and loss-of-function studies indicate that TPS1, a gene encoding trehalose-6-phosphate synthase and known to modulate glucose metabolism, activates Trk and reduces the sensitivity of yeast cells to many toxic cations. This effect is independent of known regulators of Trk, such as the Hal4 and Hal5 protein kinases and the protein phosphatase calcineurin. Mutants defective in isoform 2 of phosphoglucomutase (pgm2) and mutants defective in isoform 2 of hexokinase (hxk2) exhibit similar phenotypes of reduced Trk activity and increased sensitivity to toxic cations compared with tps1 mutants. In all cases Trk activity was positively correlated with levels of glucose phosphates (glc-1-P and glc-6-P). These results indicate that Tps1, like Pgm2 and Hxk2, increases the levels of glucose phosphates and suggest that these metabolites, directly or indirectly, activate Trk.

Adenosine Triphosphatases↗

PubMed1993

Regulation of glycolytic enzymes and the Crabtree effect in galactose-limited continuous cultures of Saccharomyces cerevisiae.

In order to determine whether the changes in the activities and mRNA levels of enzymes involved in intermediary carbon metabolism previously observed in glucose-limited continuous cultures (Sierkstra et al., 1992a) were glucose specific, we have analysed their regulation in a galactose-limited continuous culture of Saccharomyces cerevisiae. The Vmax of the galactose uptake system was shown to be dilution rate (D) dependent, comparable with the high-affinity glucose uptake. The maximum uptake was observed at D 0.2 h-1 (0.25 mmol min-1 per g) and the minimum uptake (0.1 mmol min-1 per g) at D 0.05 h-1 and 0.3 h-1. The aerobic fermentation of galactose occurred at D 0.275-0.3 h-1 which is identical to the results obtained in glucose-limited continuous cultures of this strain. Because galactose is not a repressing carbon source, this demonstrates that the Crabtree effect is not mediated by, or in any way related to glucose repression. Moreover, invertase and hexokinase I mRNA levels (both subject to glucose repression at the transcriptional level) were present when the yeast produced ethanol in galactose- and glucose-limited continuous cultures. In glucose-limited continuous cultures a decrease in alcohol dehydrogenase (I and II) mRNA levels and activity and phosphoglucomutase activity was observed with increasing dilution rates. In addition, at D 0.3 h-1, when the yeast produced ethanol, glucose-6-phosphate dehydrogenase and pyruvate decarboxylase were induced and a decrease in respiration was observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Aerobiosis↗