Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “GLYCOGEN”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 667 records · Page 37Linked to original sources
PubMed1994

Interactions between cAMP-dependent and SNF1 protein kinases in the control of glycogen accumulation in Saccharomyces cerevisiae.

The synthesis of glycogen in Saccharomyces cerevisiae is stimulated by nutrient limitation and requires both glycogen synthase and the glycogen branching enzyme. Of the two glycogen synthase genes present in yeast, GSY2 appears to be more important for the accumulation of glycogen upon entry into stationary phase. In cells grown on glucose, GSY2 mRNA levels increased approximately 10-fold during the transition from logarithmic to stationary phase. Growth of cells in glycerol, however, resulted in constitutive expression of GSY2 mRNA and the corresponding protein, GS-2, suggestive of glucose repression of GSY2. Mutants defective in the SNF1 gene, which encodes a protein kinase important in glucose repression mechanisms, are known not to accumulate glycogen. A modest 2-4-fold decrease in total GS-2 level was observed, and upon entry into stationary phase, the enzyme was blocked in the inactive, phosphorylated state in snf1 strains. The GS-2 protein is thought to be regulated by covalent phosphorylation of three COOH-terminal sites (Hardy, T.A., and Roach, P.J. (1993) J. Biol. Chem. 268, 23799-23805), removal of which results in constitutively active glycogen synthase that bypasses phosphorylation controls. Expression of COOH-terminally truncated GS-2 in snf1 cells restored glycogen accumulation, and so we propose that the SNF1 kinase controls the phosphorylation state of GS-2. Cyclic AMP pathways also exert control over glycogen accumulation. In bcy1 cells, which have constitutively active cyclic AMP-dependent protein kinase, greatly reduced levels of both GS-2 message and protein were observed. With wild type GSY2 placed under control of the ADH1 promoter, bcy1 cells did not accumulate glycogen despite increased GS-2. Overexpression of truncated GS-2, however, resulted in definite though reduced glycogen accumulation; the glycogen synthesized was structurally distinct from wild type with properties characteristic of less branched polysaccharide. We conclude that the cAMP pathway controls both the expression and the phosphorylation state of GS-2. Furthermore, other factor(s) necessary for glycogen biosynthesis, such as the branching enzyme GLC3, must also be under negative control by the cAMP pathway. The results demonstrate interactive controls of GS-2 by the cAMP-dependent and SNF1 protein kinases.

Alcohol Dehydrogenase↗
PubMed2002

New perspectives on the storage and organization of muscle glycogen.

Due to its large mass, skeletal muscle contains the largest depot of stored carbohydrate in the body in the form of muscle glycogen. Readily visualized by the electron microscope, glycogen granules appear as bead-like structures localized to specific subcellular locales. Each glycogen granule is a functional unit, not only containing carbohydrate, but also enzymes and other proteins needed for its metabolism. These proteins are not static, but rather associate and dissociate depending on the carbohydrate balance in the muscle. This review examines glycogen-associated proteins, their interactions, and roles in regulating glycogen metabolism. While certain enzymes such as glycogen synthase and glycogen phosphorylase have been extensively studied, other proteins such as the glycogen initiating and targeting proteins are just beginning to be understood. Two metabolically distinct forms of glycogen, pro- and marcoglycogen have been identified that vary in their carbohydrate complement per molecule and have different sensitivities to glycogen synthesis and degradation. Glycogen regulation takes place not only by allosteric regulation of enzymes, but also due to other factors such as subcellular location, granule size, and association with various glycogen-related proteins.

1,4-alpha-Glucan Branching Enzyme↗
PubMed1982

Development of the glycogen body through the whole length of the chick spinal cord.

The development of the glycogen body was studied throughout the entire length of the chick spinal cord. The glycogen body cells first appeared at stage 31 on each side of the ependymal septum from the 26th to 28th segments of the spinal cord. by stage 34 the paired primordia of the glycogen body extended from the 25th to 29th segments. In the middle of the structure described classically as the glycogen body (i.e., the portion restricted to the level of the spinal nerves 26-29), these primordia were fused dorsally at stage 34 and had fused completely by stage 37. The paired primordia extended from the cervical enlargement to the lumbosacral portion of the spinal cord by stage 36 and extended to the upper cervical segments by stage 38. They were totally fused throughout the entire length of the spinal cord by stage 42. The glycogen-containin cells, in the classical glycogen body level, appeared ventrolateral to the central canal at stage 35. Thereafter they increased in number and glycogen content, and extended rostrad and caudad from the classical glycogen body level. They fused to each other and then fused with the glycogen body. Therefore, the bilateral clusters of the glycogen containing cells are considered as the ventral paired primordia of the glycogen body. The development of the glycogen body is essentially the same pattern as in the classical glycogen body throughout the entire length of the spinal cord.

Animals↗
PubMed1994

Absence of glycogen cycling in cultured rat hepatocytes.

Glycogen synthesis and degradation were studied in cultured rat hepatocytes prelabeled by incubation with [14C]glucose or [14C]galactose. During prelabeling about 75% of the accumulated glycogen was synthesized from glucose and about 25% from gluconeogenic precursors. Following the labeling period, glycogen synthesis and degradation were estimated at 5 and 12.5 mM glucose and varying concentrations of insulin and glucagon. At 12.5 mM glucose and 10 nM insulin the accumulation of glycogen was comparable to in vivo values, whereas the level of radioactivity in prelabeled glycogen remained constant. Further addition of 0.1 nM glucagon resulted in constant values of both content and radioactivity of glycogen. Increasing the concentration of glucagon to 10 nM resulted in a parallel decrease of content and radioactivity in glycogen. At 5 mM glucose, 10 nM insulin, and 0.1 nM glucagon both the content and the radioactivity of glycogen were constant, whereas addition of 10 nM glucagon resulted in a parallel decrease of content and radioactivity of glycogen, which was 64% higher than that observed with 12.5 mM glucose. In the absence of insulin, prostaglandin D2 had effects similar to those of 10 nM glucagon, whereas no effects was observed in the presence of insulin. From these results and from calculated rates of glucose 6-phosphate formation, it is concluded that the rate of glycogen degradation is less than 10% of the rate of synthesis under conditions favoring glycogen accumulation. At conditions favoring glycogen degradation (10 nM insulin plus 10 nM glucagon or prostaglandin in the absence of insulin) no synthesis could be detected. Results from cells prelabeled with [14C]galactose suggested that glycogen degradation is not an absolutely ordered process, but that some random degradation takes place.

Animals↗
PubMed1998

Training decreases muscle glycogen turnover during exercise.

The present study was undertaken to determine the effects of endurance training on glycogen kinetics during exercise. A new model describing glycogen kinetics was applied to quantitate the rates of synthesis and degradation of glycogen. Trained and untrained rats were infused with a 25% glucose solution with 6-3H-glucose and U-14C-lactate at 1.5 and 0.5 microCi x min(-1) (where 1 Ci=3.7 x 10(10) Bq), respectively, during rest (30 min) and exercise (60 min). Blood samples were taken at 10-min intervals starting just prior to isotopic infusion, until the cessation of exercise. Tissues harvested after the cessation of exercise were muscle (soleus, deep, and superficial vastus lateralis, gastrocnemius), liver, and heart. Tissue glycogen was quantitated and analyzed for incorporation of 3H and 14C via liquid scintillation counting. There were no net decreases in muscle glycogen concentration from trained rats, whereas muscle glycogen concentration decreased to as much as 64% (P < 0.05) in soleus in muscles from untrained rats after exercise. Liver glycogen decreased in both trained (30%) and untrained (40%) rats. Glycogen specific activity increased in all tissues after exercise indicating isotope incorporation and, thus, glycogen synthesis during exercise. There were no differences in muscle glycogen synthesis rates between trained and untrained rats after exercise. However, training decreased muscle glycogen degradation rates in total muscle (i.e., the sum of the degradation rates of all of the muscles sampled) tenfold (P < 0.05). We have applied a model to describe glycogen kinetics in relation to glucose and lactate metabolism during exercise in trained and untrained rats. Training significantly decreases muscle glycogen degradation rates during exercise.

Animals↗
PubMed1983

Acetaminophen-induced hepatic glycogen depletion and hyperglycemia in mice.

Two hours following administration of a hepatotoxic dose of acetaminophen (500 mg/kg, i.p.) to mice, liver sections stained with periodic acid Schiff reagent showed centrilobular hepatic glycogen depletion. A chemical assay revealed that following acetaminophen administration (500 mg/kg) hepatic glycogen was depleted by 65% at 1 hr and 80% at 2 hr, whereas glutathione was depleted by 65% at 0.5 hr and 80% at 1.5 hr. Maximal glycogen depletion (85% at 2.5 hr correlated with maximal hyperglycemia (267 mg/100 ml at 2.5 hr). At 4.0 hr following acetaminophen administration, blood glucose levels were not significantly different from saline-treated animals; however, glycogen levels were still maximally depleted. A comparison of the dose-response curves for hepatic glycogen depletion and glutathione depletion showed that acetaminophen (50-500 mg/kg at 2.5 hr) depleted both glycogen and glutathione by similar percentages at each dose. Since acetaminophen (100 mg/kg at 2.5 hr) depleted glutathione and glycogen by approximately 30%, evidence for hepatotoxicity was examined at this dose to determine the potential importance of hepatic necrosis in glycogen depletion. Twenty-four hours following administration of acetaminophen (100 mg/kg) to mice, histological evidence of hepatic necrosis was not detected and serum glutamate pyruvate transaminase (SGPT) levels were not significantly different from saline-treated mice. The potential role of glycogen depletion in altering the acetaminophen-induced hepatotoxicity was examined subsequently. When mice were fasted overnight, hepatic glutathione and glycogen were decreased by 40 and 75%, respectively, and fasted animals showed a dramatic increase in susceptibility to acetaminophen-induced hepatotoxicity as measured by increased SGPT levels. Availability of glucose in the drinking water (5%) overnight resulted in glycogen levels similar to those in fed animals, whereas hepatic glutathione levels were not significantly different from those of fasted animals. Fasted animals and animals given glucose water overnight were equally susceptible to acetaminophen-induced hepatotoxicity, as quantitated by increases in SGPT levels 24 hr after drug administration. The potential role of a reactive metabolite in glycogen depletion was investigated by treating mice with N-acetylcysteine to increase detoxification of the reactive metabolite. N-Acetylcysteine treatment of mice prevented acetaminophen-induced glycogen depletion.

Acetaminophen↗
PubMed1987

Differences between glycogen synthases from rat and rabbit skeletal muscle as indicated by phosphopeptide maps.

Glycogen synthase I was purified from rat skeletal muscle. On sodium dodecyl sulfate polyacrylamide gel electrophoresis, the enzyme migrated as a major band with a subunit Mr of 85,000. The specific activity (24 units/mg protein), activity ratio (the activity in the absence of glucose-6-P divided by the activity in the presence of glucose-6-P X 100) (92 +/- 2) and phosphate content (0.6 mol/mol subunit) were similar to the enzyme from rabbit skeletal muscle. Phosphorylation and inactivation of rat muscle glycogen synthase by casein kinase I, casein kinase II (glycogen synthase kinase 5), glycogen synthase kinase 3 (kinase FA), glycogen synthase kinase 4, phosphorylase b kinase, and the catalytic subunit of cAMP-dependent protein kinase were similar to those reported for rabbit muscle synthase. The greatest decrease in rat muscle glycogen synthase activity was seen after phosphorylation of the synthase by casein kinase I. Phosphopeptide maps of glycogen synthase were obtained by digesting the different 32P-labeled forms of glycogen synthase by CNBr, trypsin, or chymotrypsin. The CNBr peptides were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and the tryptic and chymotryptic peptides were separated by reversed-phase HPLC. Although the rat and rabbit forms of synthase gave similar peptide maps, there were significant differences between the phosphopeptides derived from the N-terminal region of rabbit glycogen synthase and the corresponding peptides presumably derived from the N-terminal region of rat glycogen synthase. For CNBr peptides, the apparent Mr was 12,500 for rat and 12,000 for the rabbit. The tryptic peptides obtained from the two species had different retention times. A single chymotryptic peptide was produced from rat skeletal muscle glycogen synthase after phosphorylation by phosphorylase kinase whereas two peptides were obtained with the rabbit enzyme. These results indicate that the N-terminus of rabbit glycogen synthase, which contains four phosphorylatable residues (Kuret et al. (1985) Eur. J. Biochem. 151, 39-48), is different from the N-terminus of rat glycogen synthase.

Animals↗
PubMed1976

Glycogen degradation during migration of presumptive cell types in Dictyostelium discoideum.

During the time course of differentiation in Dictyostelium discoideum, glycogen was found to accumulate from the amoebae stage to the culmination stage of development. Upon sorocarp formation (23 h), glycogen was rapidly degraded. Ultramicrotechniques, utilizing amplification of glycogen by enzymatic cycling, were used to follow glycogen metabolism in pre-stalk and prespore cells during the differentiation cycle. Both cell types accumulated glycogen at nearly the same rate. By the pseudoplasmodium stage of development glycogen had accumulated to 50% of its maximum value, and no differences were found between pre-stalk and pre-spore cells. Glycogen was degraded as pre-stalk cells migrated into the position for stalk construction. At the culmination stage of development stalk cells near the base were devoid of glycogen while pre-stalk cells near the apex of the stalk showed no loss of glycogen. The complete loss of glycogen from stalk cells occurred over a distance occupied by approximately 100 cells, and over a time period of approx. 1 h. Pre-spore cells at the culmination stage showed no loss of glycogen even though separated from stalk cells by only a thin cellulose sheath. The degradation of prespore cell glycogen did not commence until stalk construction was completed and the pre-spore mass had reached the apex of the stalk. Pre-spore cells at the culmination stage contained high levels of glycogen while only 2 h later, total degradation had occurred.

Amoeba↗
PubMed1977

A protein-bound glycogen component of rat liver.

It has long been claimed, but frequently disputed, that part of the glycogen in rat liver is insoluble in 10% trichloroacetic acid, and a physiological significance was ascribed to the existence of the two pools of glycogen, desmo-glycogen, the insoluble form, and lyo-glycogen, the soluble component. Desmo-glycogen was thought to owe its acid insolubility to a covalent binding to protein. Recent claims that glycogen, similarly insoluble in acid, can be synthesized in vitro have renewed the interest in desmo-glycogen. We have obtained trichloroacetic acid-insoluble glycogen from rat liver and find that, despite subjecting the glycogen to proteolysis, peptide material remains in close association with the glycogen through a number of purification procedures and is freed from glycogen only by enzymic decomposition of the latter. The tenacity with which the glycogen and peptide material remain in association with each other is suggestive of the occurrence of protein-bound glycogen.

Amino Acids↗
PubMed1994

Characterization of human endometrial glycogen synthase.

OBJECTIVE: To characterize and compare human endometrial glycogen synthase complementary DNA (cDNA) to the published rat liver and human skeletal muscle glycogen synthase transcripts and then to determine whether P-induced glycogen accumulation in endometrium is accompanied by a rise in glycogen synthase transcripts. DESIGN: We performed bidirectional sequencing of segments of endometrial glycogen synthase with chosen primers. Endometrial tissue was cultured up to 72 hours with and without P. Total RNA was extracted from in vivo and cultured endometrium and used as template for semiquantitative reverse transcription polymerase chain reaction. SETTING: Normal human volunteers in an academic research environment. PATIENTS: Premenopausal women with histologically normal endometrium undergoing hysterectomy. INTERVENTIONS: ENdometrium was maintained in culture with and without exposure to P. MAIN OUTCOME MEASURE: Human endometrial glycogen synthase cDNA nucleotide sequence and messenger RNA (mRNA) levels. RESULTS: In all areas sequenced, human endometrial glycogen synthase cDNA is identical to that of human skeletal muscle. Conversely, it is only 80% similar to rat and human liver glycogen synthase cDNA in these areas. Tissue levels of glycogen synthase transcript did not change in vivo or in vitro after P exposure. CONCLUSIONS: Human endometrial glycogen synthase is similar if not identical to human skeletal muscle glycogen synthase. The mechanism for P induction of endometrial glycogen does not require change in the concentration of glycogen synthase transcripts.

Adult↗
PubMed1998

Microfluorometric analyses of glycogen in freshly dissected, single skeletal muscle fibres of the cane toad using a mechanically skinned fibre preparation.

The main objective of this study was to analyse glycogen in single muscle fibres, using a recently developed microfluorometric method which detects subpicomol amounts of NADPH, glucose and glycogen (as glucosyl units) (detection limit 0.16-0.17 pmol in a 25 nl sample) without fluorochrome amplification. The fibres were freshly dissected from the twitch region of the iliofibularis muscle of the cane toad (Bufo marinus), and were mechanically skinned under paraffin oil to gain access to the intracellular compartments. The results show that (1) glycogen concentrations in toad skeletal muscle fibres range between 25.8 and 369 mmol glucosyl units/litre fibre volume; (2) there is a large variation in glycogen content between individual fibres from the iliofibularis muscle of one animal; (3) there are seasonal differences in the glycogen content of toad single muscle fibres; (4) the total amount of glycogen in single muscle fibres of the toad does not decrease significantly when storing the tissue, under paraffin oil, at 20-25 degree C for up to 6 h or at 4 degree C for up to 24 h; and (5) 15-26% of fibre glycogen can be washed in an aqueous solution at pH 5-7, within 5 min, while 74-85% of fibre glycogen remains associated with the washed skinned fibre, even after 40 min exposure of the skinned fibre preparation to the aqueous environment. The retention of most glycogen in the fibre preparation after mechanical removal of the plasma membrane and extensive washing indicates that in toad skeletal muscle fibres the largest proportion of glycogen is tightly bound to intracellular structures. The results also show that the skinned muscle fibre preparation is well suited for microfluorometric glycogen determination, since low molecular weight non-glycogen contributors to the fluorescence signal can be removed from the myoplasmic space prior to the glycogen hydrolysis step.

Animals↗
PubMed1989

Glycogen synthesis during exercise and rest with carbohydrate feeding in males and females.

Since it has been demonstrated that endurance-trained cyclists are able to synthesize glycogen during mild exercise, glycogen synthesis was investigated in non-endurance-trained males and females as well. Seven males and nine females exercised on a cycle ergometer to deplete muscle glycogen. After the exhaustive exercise and taking a muscle biopsy, the males either exercised 2.5 h at 40% of maximal work load (trial A) or rested for 2.5 h (trial B). In both trials the subjects drank a 25% maltodextrin-fructose solution. After 2.5 h of exercise or rest, a second muscle biopsy was taken for determination of glycogen and for histochemistry (ATPase and PAS). In the females glycogen synthesis was only studied during 2.5 h rest, after prior glycogen depletion. In the male subjects, during mild exercise with carbohydrate feeding muscle glycogen did not increase. During rest muscle glycogen increased in the males from 123 +/- 49 mmol/kg DW at exhaustion to 229 +/- 70 mmol/kg DW (P less than 0.001), resulting in a net increase of 42 mmol/kg DW/h. Glycogen synthesis during rest occurred both in type I and type II fibers. In the females, during 2.5 h of rest, muscle glycogen increased from 130 +/- 56 mmol/kg DW at exhaustion to 224 +/- 51 mmol/kg DW, resulting in a net increase of 37 mmol/kg DW/h. The results demonstrate that glycogen synthesis during mild exercise does not occur in non-endurance-trained athletes, whereas in the resting state glycogen synthesis in non-endurance-trained males is not different from endurance-trained cyclists. In addition, glycogen synthesis during rest is similar in males and females.

Adult↗
PubMed1996

Overexpression of muscle glycogen phosphorylase in cultured human muscle fibers causes increased glucose consumption and nonoxidative disposal.

The effect of increased expression of glycogen phosphorylase on glucose metabolism in human muscle was examined in primary cultured fibers transduced with recombinant adenovirus AdCMV-MGP encoding muscle glycogen phosphorylase. Increments of 20-fold in total enzyme activity and of 14-fold of the active form of the enzyme were associated with a 30% reduction in basal glycogen levels. Total glycogen synthase activity was doubled in AdCMV-MGP-transduced cells even though the activity ratio was decreased. Incubation with forskolin, which inactivated glycogen synthase and activated glycogen phosphorylase, induced greater net glycogenolysis in engineered cells. In unstimulated fibers, lactate production was three times higher in AdCMV-MGP fibers as compared with controls, despite similar rates of glycogenolysis. In transduced fibers incubated with 2-deoxyglucose, the level of 2-deoxyglucose 6-phosphate was about 8-fold elevated over the control even though hexokinase activity was unmodified in AdCMV-MGP fibers. Overexpression of glycogen phosphorylase also led to enhancement of [U-14C]glucose incorporation into glycogen, lactate, and lipid. Accordingly, determination of lipid cell content revealed that engineered cells were accumulating lipids. Furthermore, 14CO2 formation from [U-14C]glucose was 1.6-fold higher, whereas 14CO2 formation from [6-14C]glucose was unmodified, in AdCMV-MGP fibers. Our data show that in human skeletal muscle cells in culture, the increase in glycogen phosphorylase activity is able to up-regulate glycogen synthase activity indicating the enhancement of glycogen turnover. We suggest that the increase in glycogen phosphorylase and, thereby, in glycogen metabolism, is sufficient to enhance glucose uptake in the muscle cell. Glucose taken up by engineered muscle cells is essentially disposed of through nonoxidative metabolism and converted into lactate and lipid.

Carbon Radioisotopes↗
PubMed1961

Glycogen synthesis from uridine diphosphate glucose. The distribution of the enzyme in liver cell fractions.

The distribution in liver cell fractions of UDPG-glycogen transferase has been studied. In fasting animals which have been refed 6 hours before sacrifice, the distribution of the enzyme in the various cell fractions can be correlated with the glycogen content of each fraction. A purified glycogen fraction has been prepared by differential centrifugation in sucrose gradients. This glycogen fraction contains vesicular structures which resemble those seen in association with glycogen deposits in the intact liver cell. In addition, the glycogen pellet contains UDPG-glycogen transferase in high specific activity. Subfractionation of the glycogen pellet separates the majority of vesicular elements from the bulk of transferase activity and glycogen. The evidence presented suggests that the presence of UPDG-glycogen transferase in the glycogen pellet is to be attributed to its binding to glycogen rather than to its association with the structural elements found in the glycogen fraction.

Animals↗
PubMed1990

Lysosomal glycogen accumulation in rat liver and its in vivo kinetics after a single intraperitoneal injection of acarbose, an alpha-glucosidase inhibitor.

A single intraperitoneal injection of acarbose (400 mg/kg) into rats caused lysosomal accumulation of glycogen in the liver, mimicking the cytological characteristics of human glycogen storage disease type II (Pompe's disease). The animal model is therefore useful for studying the pathogenesis of the disease. In the present study, we applied this model to examine the lysosomal hydrolytic pathway of glycogen in vivo. To quantify the lysosomal glycogen, the lysosome-rich fraction was rapidly prepared from liver homogenate by agglutination in the presence of Ca2+. Then the fraction was treated with alpha-amylase in isotonic medium to remove cytosolic glycogen, followed by transfer to hypotonic conditions in the presence of Triton X-100 to destroy total glycogen. The amount of lysosomal glycogen was calculated from the difference between the glycogen levels measured before and after the treatment under hypotonic conditions, and then it was corrected based on measurements of the intactness (%) of lysosomes and the recovery (%) of the lysosomal marker enzyme (beta NAGase). We observed no measurable lysosomal glycogen in normal liver by this method, and this was confirmed by electron microscopy. After administration of acarbose, the lysosomal glycogen level increased to 2.5 mg/g liver within 2 days, and then decreased gradually at a rate of 0.4 mg/day/g. The accumulation of glycogen in the lysosomes at an initial velocity of 1.5 mg/day/g liver may be considered as the amount of glycogen that would normally be degraded by acid alpha-glucosidase. Therefore, assuming that the liver breaks down about 40 mg glycogen/day/g, we estimated that about 3% of the glycogen would be hydrolyzed by the lysosomal pathway.

Acarbose↗
PubMed1983

The calmodulin-dependent glycogen synthase kinase from rabbit skeletal muscle. Purification, subunit structure and substrate specificity.

A calmodulin-dependent glycogen synthase kinase distinct from phosphorylase kinase has been purified approximately equal to 5000-fold from rabbit skeletal muscle by a procedure involving fractionation with ammonium sulphate (0-33%), and chromatographies on phosphocellulose, calmodulin-Sepharose and DEAE-Sepharose. 0.75 mg of protein was obtained from 5000 g of muscle within 4 days, corresponding to a yield of approximately equal to 3%. The Km for glycogen synthase was 3.0 microM and the V 1.6-2.0 mumol min-1 mg-1. The purified enzyme showed a major protein staining band (Mr 58 000) and a minor component (Mr 54 000) when examined by dodecyl sulphate polyacrylamide gel electrophoresis. The molecular weight of the native enzyme was determined to be 696 000 by sedimentation equilibrium centrifugation, indicating a dodecameric structure. Electron microscopy suggested that the 12 subunits were arranged as two hexameric rings stacked one upon the other. Following incubation with Mg-ATP and Ca2+-calmodulin, the purified protein kinase underwent an 'autophosphorylation reaction'. The reaction reached a plateau when approximately equal to 5 mol of phosphate had been incorporated per 58 000-Mr subunit. Both the 58 000-Mr and 54 000-Mr species were phosphorylated to a similar extent. Autophosphorylation did not affect the catalytic activity. The calmodulin-dependent protein kinase initially phosphorylated glycogen synthase at site-2, followed by a slower phosphorylation of site-1 b. The protein kinase also phosphorylated smooth muscle myosin light chains, histone H1, acetyl-CoA carboxylase and ATP-citrate lyase. These findings suggest that the calmodulin-dependent glycogen synthase kinase may be a enzyme of broad specificity in vivo. Glycogen synthase kinase-4 is an enzyme that resembles the calmodulin-dependent glycogen synthase kinase in phosphorylating glycogen synthase (at site-2), but not glycogen phosphorylase. Glycogen synthase kinase-4 was unable to phosphorylate any of the other proteins phosphorylated by the calmodulin-dependent glycogen synthase kinase, nor could it phosphorylate site 1 b of glycogen synthase. The results demonstrate that glycogen synthase kinase-4 is not a proteolytic fragment of the calmodulin-dependent glycogen synthase kinase, that has lost its ability to be regulated by Ca2+-calmodulin.

Animals↗
PubMedDate not supplied

Blood glycogen and metabolic control in diabetes mellitus.

The relationship between metabolic control and leukocyte glycogen content in diabetes mellitus was re-evaluated, blood glycogen being measured by an enzymatic procedure. In 30 healthy subjects, fasting blood glycogen averaged 50.6 +/- 2.8 mg l-1 or 7.45 +/- 0.42 ng 10(3)-cells-1, the latter value being unaffected during a 60-min period of induced hyperglycaemia. Comparable levels were found in 18 Type 1 insulin-treated diabetic patients (blood glycogen 50.4 +/- 4.6 mg l-1, leukocyte glycogen 6.92 +/- 0.50 ng 10(3)-cells-1), 6 insulin-treated diabetic patients presenting with chronic pancreatitis (blood glycogen 62.2 +/- 9.3 mg l-1, leukocyte glycogen 6.69 +/- 0.70 ng 10(3)-cells-1) and 12 Type 2 insulin-treated patients (blood glycogen 53.7 +/- 4.3 mg l-1, leukocyte glycogen 7.51 +/- 0.44 ng 10(3)-cells-1). In severely ketotic patients, leukocyte counts and blood glycogen (160.8 +/- 29.6 mg l-1, p less than 0.01 vs stable diabetic patients) were increased, but the leukocytic glycogen content was not significantly affected either before or during intensive insulin therapy and rehydration. The leukocyte glycogen content was abnormally low, however, in 9 untreated Type 2 diabetic patients (5.29 +/- 0.39 ng 10(3)-cells-1, p less than 0.02 vs healthy subjects) and abnormally high (10.77 +/- 0.65 ng 10(3)-cells-1, p less than 0.005 vs healthy individuals) in 30 Type 2 patients treated by sulphonylurea, alone or in combination with insulin. No correlation was found between leukocyte glycogen and either fasting plasma glucose or HbA1c.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1990

Glucose uptake and transport in contracting, perfused rat muscle with different pre-contraction glycogen concentrations.

1. Glucose uptake and transport, muscle glycogen, free glucose and glucose-6-phosphate concentrations were studied in perfused resting and contracting rat skeletal muscle with different pre-contraction glycogen concentrations. Rats were pre-conditioned by a combination of swimming exercise and diet, resulting in either low (glycogen-depleted rats), normal (control rats) or high (supercompensated rats) muscle glycogen concentrations at the time their hindlimbs were perfused. 2. Compared with control rats, pre-contraction muscle glycogen concentration was approximately 40% lower in glycogen-depleted rats, whereas it was 40% higher in supercompensated rats. Muscle glycogen break-down correlated positively (r = 0.76; P less than 0.001) with pre-contraction muscle glycogen concentration. 3. Glucose uptake during contractions was approximately 50% higher in glycogen-depleted hindquarters than in control hindquarters; in supercompensated hindquarters it was 30% lower. When rats with similar muscle glycogen concentrations were compared, glucose uptake in hindquarters from rats that had exercised on the preceding day was approximately 20% higher than in hindquarters from rats that had not exercised on the preceding day. 4. Muscle membrane glucose transport, as measured by the rate of accumulation of 14C-3-O-methylglucose in the contracting muscles, was 25% lower in supercompensated than in glycogen-depleted muscles at the onset as well as at the end of the 15 min contraction period. 5. Intracellular concentrations of free glucose and glucose-6-phosphate were higher at rest and during the entire 15-min stimulation period in supercompensated muscles than in glycogen-depleted muscles, and glucose uptake during contractions correlated negatively with free glucose (r = -0.52; P less than 0.01) as well as with glucose-6-phosphate (r = -0.49; P less than 0.01) concentrations. 6. It is concluded that: (a) The rate of glucose uptake in contracting skeletal muscle is dependent on the pre-contraction muscle glycogen concentration. Regulating mechanisms include limitations of membrane glucose transport as well as of glucose metabolism. (b) Contractions on the preceding day have a stimulating effect on glucose uptake during contractions of the same muscles on the next day.

3-O-Methylglucose↗