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B Crabtree

Publications and source records attributed to B Crabtree.

At least 37 records · Page 2Linked to original sources
PubMed1987

Measurement of the rate of substrate cycling between acetate and acetyl-CoA in sheep muscle in vivo. Effects of infusion of acetate.

1. The rate of substrate cycling between acetate and acetyl-CoA in sheep hindlimb muscle was measured from the decrease of [14C]acetate specific radioactivity across the tissue in vivo. 2. The rate of cycling was 0.005-0.015 mumol/min per g and, in two of the three sheep investigated, was approximately doubled as a result of infusing 2 mol acetate/day intravenously. 3. The results indicate that this cycle contributes only approx. 0.5% of the total heat produced by the animal and would probably make only a relatively small contribution to any heat increment of feeding associated with diets producing large amounts of acetate.

Acetates↗
PubMed1987

Hormonal regulation of the rate of the glycogen/glucose-1-phosphate cycle in skeletal muscle.

A method is presented which allows simultaneous estimation of rates of glycogen synthesis and glycogenolysis in an isolated incubated skeletal muscle, thus allowing measurement of the glycogen/glucose-1-phosphate substrate cycling rate. In the absence of hormonal additions, the measured rates of glycogen synthesis and breakdown were similar [respectively, 0.54 +/- 0.12 (8) and 0.74 +/- 0.10 (8) mumol glucosyl equiv. h-1 (g tissue)-1]. Incremental addition of insulin stimulated glycogen synthesis up to threefold and reduced glycogenolysis by about sevenfold; the half-maximally effective concentration of insulin on both processes was about 100 microU/ml (0.7 nM). Incremental addition of adrenaline (in the presence of 1 mU insulin/ml) caused a dramatic increase in the glycogenolytic rate (about 15-fold), but a much less marked inhibition of glycogen synthetic rate. In addition to hormonal manipulation of the muscle preparation in vitro, the effects of cold exposure, the hyperthyroid state, a single exercise bout and exercise-training of animals in vivo on the rates of glycogen synthesis and breakdown in the isolated incubated muscle preparation have been investigated. Significant changes in measured glycogen synthesis, breakdown and glycogen/glucose-1-phosphate cycling have been observed, both under basal conditions and in response to hormonal additions in vitro. The results are discussed with respect to the possible physiological importance of this substrate cycle.

Adaptation, Physiological↗
PubMed1985

A metabolic switch produced by enzymically interconvertible forms of an enzyme.

A system of enzymically interconvertible enzyme forms (interconversion cycle) is known to increase the sensitivity to regulators. Here it is shown that this system may also provide a discrete switching mechanism in which the response is maintained when the stimulus is removed. These switches may be important in producing discontinuous metabolic changes such as those associated with cell differentiation.

Cell Differentiation↗
PubMed1985

The role of high rates of glycolysis and glutamine utilization in rapidly dividing cells.

The rates of utilization of both glucose and glutamine are high in rapidly dividing cells such as enterocytes, lymphocytes, thymocytes, tumour cells; the oxidation of both glucose and glutamine is only partial, glucose to lactate and glutamine to glutamate, alanine or aspartate; and these partial processes are termed glycolysis and glutaminolysis respectively. Both processes generate energy and also provide precursors for important biosynthetic processes in such cells. However, the rates of utilization of precursors for macromolecular biosynthesis are very low in comparison to the rates of partial oxidation, and energy generation per se may not be the correct explanation for high rates of glycolysis and glutaminolysis in these cells since oxidation is only partial and other fuels can be used to generate energy. Both the high fluxes and the metabolic characteristics of these two processes can be explained by application of quantitative principles of control as applied to branched metabolic pathways (Crabtree & Newsholme, 1985). If the flux through one branch is greatly in excess of the other, then the sensitivity of the flux of the low-flux pathway to regulators is very high. Hence, it is suggested that, in rapidly dividing cells, high rates of glycolysis and glutaminolysis are required not for energy or precursor provision per se but for high sensitivity of the pathways involved in the use of precursors for macromolecular synthesis to specific regulators to permit high rates of proliferation when required - for example, in lymphocytes in response to a massive infection.

Animals↗
PubMed1985

Glutamine metabolism in lymphocytes: its biochemical, physiological and clinical importance.

Glutamine is utilized at a high rate (fourfold higher than that of glucose) by isolated incubated lymphocytes and produces glutamate, aspartate, lactate and ammonia. The pathway for glutamine metabolism includes the reactions catalysed by glutaminase, aspartate aminotransferase, oxoglutarate dehydrogenase, succinate dehydrogenase, fumarase, malate dehydrogenase and phosphoenolpyruvate carboxykinase. In fact little if any of the carbon of the glutamine that is used is converted to acetyl-CoA for complete oxidation. For this reason, the oxidation of glutamine is only partial and, in an analogous manner to the terminology used to describe the partial oxidation of glucose to lactate as glycolysis, the term glutaminolysis is used to describe the process of partial glutamine oxidation. The role of glutaminolysis in lymphocytes and perhaps other rapidly dividing cells is to provide both nitrogen and carbon for precursors for synthesis of macromolecules (e.g. purines and pyrimidines for DNA and RNA) and also energy. However, the rate of glutamine utilization by lymphocytes is markedly in excess of the precursor requirements (which are at most 4%) and if glutamine was vitally important in energy production it would be expected that more would be converted to acetyl-CoA for complete oxidation via the Krebs cycle. Indeed most of the energy for lymphocytes may be obtained by the complete oxidation of fatty acids and ketone bodies. Consequently the role of the high rate of glutaminolysis in lymphocytes and other rapidly dividing cells may be identical to that of glycolysis: the high rates provide ideal conditions for the precise and sensitive control of the rate of use of the intermediates of these pathways for biosynthesis when required. High rates of glycolysis and glutaminolysis can be seen as part of a mechanism of control to permit synthesis of macromolecules when required without any need for extracellular signals to make more glucose or glutamine available for these cells. In order to maintain a high rate of glutaminolysis despite fluctuation in the plasma level of glutamine, the flux through the glutaminolytic pathway can be controlled and the key processes in the lymphocyte that may play a role in this process include glutamine transport across the cell and mitochondrial membranes, glutaminase and oxoglutarate dehydrogenase. Changes in the intracellular concentration of Ca2+ may play a role in control of one or more of these reactions.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acids↗
PubMed1984

Measurement of the rate of substrate cycling between fructose 6-phosphate and fructose 1,6-bisphosphate in skeletal muscle by using a single-isotope technique.

The effects of several agents on the rates of the fructose 6-phosphate/fructose 1,6-bisphosphate substrate cycle were measured in incubated epitrochlearis muscles of the rat by monitoring the transfer of radiolabel from [6-14C]glucose to the 1-position of glucose residues in glycogen. The cycling rates observed were almost identical with those previously obtained by using the well-established dual-isotope technique. In particular, it was found that the beta-adrenoceptor agonist isoprenaline increased the cycling rate about 12-fold.

Animals↗
PubMed1981

The activities and intracellular distributions of enzymes of carbohydrate, lipid and ketone-body metabolism in lactating mammary glands from ruminants and non-ruminants.

1. The activities of several enzymes of carbohydrate, lipid, acetate and ketone-body metabolism were measured in lactating mammary glands from rats, mice, rabbits, guinea pigs, sows, sheep, cows and goats. The intracellular distributions of many of the enzymes were measured by fractional extraction. 2. Acetyl-CoA synthetase was predominantly cytoplasmic in rats and guinea pigs, but was more mitochondrial in the other species. The different location of this enzyme in rats and mice is discussed in relation to the disposal of reducing equivalents. 3. 3-Oxo acid CoA-transferase and acetoacetyl-CoA thiolase assayed at 600 microM-CoA were predominantly mitochondrial in all species investigated. Acetoacetyl-CoA thiolase assayed at 8 microM-CoA was predominantly cytoplasmic, except in rabbits and guinea pigs. Ruminants appeared to possess little, if any, of the cytoplasmic enzyme. 4. The activities and distributions of NADP-isocitrate dehydrogenase were consistent with a role in supplying cytoplasmic NADPH in ruminant tissue, and indicated that this system may also occur in guinea pigs.

Acetates↗
PubMed1979

Use of enzyme activities as indices of maximum rates of fuel utilization.

It can be shown theoretically and experimentally that the maximum activities in vitro of enzymes that catalyse near-equilibrium reactions in vivo must be considerably higher than the maximum flux through that pathway. Consequently, the activities of such enzymes cannot provide quantitative information on the maximum possible flux through a pathway. On the other hand, the maximum activity of an enzyme that catalyses a non-equilibrium reaction in vivo may provide quantitative information. Such possibilities must be tested experimentally. Thus the maximum flux through a given metabolic pathway is measured (or calculated) and compared with the maximum in vitro activities of enzymes that catalyse non-equilibrium reactions in that pathway. Catalytic activities similar to the flux suggest that such enzymes may be useful as flux indicators. For example, phosphorylase or phosphofructokinase activities provide a quantitative indication of maximum flux through glycolysis-from-glycogen (i.e. anaerobic glycolysis); hexokinase activities provide a quantitative indication of maximum flux through glycolysis-from-glucose; 2-oxoglutarate dehydrogenase activities provide a quantitative indication of maximum flux through the citric acid cycle. The advamtages of the use of enzyme activities in this manner include simplicity, general applicability to pathways, tissues and animals, and minimum intervention (particularly in larger animals including the human species). One disadvantage is that the properties of the enzyme must be known in detail before an assay that gives maximum activities can be developed, and the properties of enzymes that catalyse non-equilibrium reactions may be complex. These considerations emphasize the dangers of quantitative interpretation of the maximum flux through pathways from 'near-equilibrium' enzymes or from 'non-equilibrium' enzymes whose properties have been inadequately studied.

Adenosine Triphosphate↗
PubMed1977

The activities of 2-oxoglutarate dehydrogenase and pyruvate dehydrogenase in hearts and mammary glands from ruminants and non-ruminants.

1. The activities of 2-oxoglutarate dehydrogenase (EC 1.2.4.2) were measured in hearts and mammary glands of rats, mice, rabbits, guinea pigs, cows, sheep, goats and in the flight muscles of several Hymenoptera. 2. The activity of 2-oxoglutarate dehydrogenase was similar to the maximum flux through the tricarboxylic acid cycle in vivo. Therefore measuring the activity of this enzyme may provide a simple method for estimating the maximum flux through the cycle for comparative investigations. 3. The activities of pyruvate dehydrogenase (EC 1.2.4.1) in mammalian hearts were similar to those of 2-oxoglutarate dehydrogenase, suggesting that in these tissues the tricarboxylic acid cycle can be supplied (under some conditions) by acetyl-CoA derived from pyruvate alone. 4. In the lactating mammary glands of the rat and mouse, the activities of pyruvate dehydrogenase exceeded those of 2-oxoglutarate dehydrogenase, reflecting a flux of pyruvate to acetyl-CoA for fatty acid synthesis in addition to that of oxidation via the tricarboxylic acid cycle. In ruminant mammary glands the activities of pyruvate dehydrogenase were similar to those of 2-oxoglutarate dehydrogenase, reflecting the absence of a significant flux of pyruvate to fatty acids in these tissues.

Acetyl Coenzyme A↗