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1.
W.J. Malaisse F. Malaisse-Lagae A. Sener 《Biochimica et Biophysica Acta (BBA)/General Subjects》1984,797(2):194-202
1. Because L-asparagine augments insulin release evoked by L-leucine, the metabolism of these two amino acids was investigated in rat pancreatic islets. 2. L-Leucine inhibited the uptake and deamidation of L-asparagine, but failed to exert any obvious primary effect upon the further catabolism of aspartate derived from exogenous asparagine. 3. L-Asparagine augmented the oxidation of L-leucine, and effect possibly attributable to activaion of 2-ketoisocaproate dehydrogenase. 4. The association of L-asparagine and L-leucine exerted a sparing action on the utilization of endogenous amino acids, so that the integrated rate of nutrients oxidation was virtually identical in the sole presence of L-leucine and simultaneous presence of L-asparagine and L-leucine, respectively. 5. It is proposed that the enhancing action of L-asparagine upon insulin release evoked by L-leucine is attributable to an increased generation rate of cytosolic NADPH rather than any increase in nutrients oxidation. 相似文献
2.
Stimulation by D-glucose of mitochondrial oxidative events in islet cells. 总被引:1,自引:0,他引:1 下载免费PDF全文
Antibodies were elicited to FAD by using the hapten N-6-(6-aminohexyl)-FAD conjugated to the immunogenic carrier protein bovine serum albumin. Cross-reactivity was determined by Ouchterlony double-diffusion analysis with N-6-(6-aminohexyl)-FAD coupled to rabbit serum albumin. Anti-FAD IgG was partially purified by (NH4)2SO4 precipitation followed by DEAE-cellulose/CM-cellulose and bovine serum albumin-agarose chromatography. The partially purified anti-FAD IgG fraction failed to inhibit the catalytic activities of the flavin-containing enzymes nitrate reductase, xanthine oxidase and succinate dehydrogenase, whereas enzyme activity could be inhibited by addition of antibodies elicited against the native proteins. However, the partially purified anti-FAD IgG fraction could be used as a highly sensitive and specific probe to detect proteins containing only covalently bound flavin, such as succinate dehydrogenase, p-cresol methylhydroxylase and monoamine oxidase, by immuno-blotting techniques. Detection limits were estimated to be of the order of femtomolar concentrations of FAD with increased sensitivity for the 8 alpha-N(3)-histidyl linkage compared with 8 alpha-O-tyrosyl substitution. 相似文献
3.
In tumoral islet cells (RINm5F line) the phosphorylation of D-fructose is catalyzed by hexokinase rather than fructokinase. Fructose 6-phosphate appears to be preferentially channelled into the pentose cycle, as suggested by a ratio of D-[1-14C]fructose/D-[U-14C]fructose oxidation close to 2.7, the failure to generate 14C-labelled lactate from D-[1-14C]fructose and a poor metabolic response to menadione. When the islet cells are exposed to both D-fructose and D-glucose, however, the metabolism of the former hexose is dramatically modified, fructose 6-phosphate being now formed at a lower rate and preferentially channelled into the glycolytic pathway. These findings illustrate the existence of regulatory steps in fructose catabolism located distally to its site of phosphorylation. 相似文献
4.
5.
Greta Marynissen Abdullah Sener Willy J. Malaisse 《Molecular and cellular biochemistry》1992,113(2):105-121
A multifactorial quantitative analysis of oscillations in glycolysis was conducted in the postmicrosomal supernatant of rat muscle homogenates incubated in the presence of yeast hexokinase. Oscillations in adenine nucleotides, D-fructose 1,6-bisphosphate, triose phosphates, L-glycerol 3-phosphate, 3HOH generation from D-[5-3H]glucose, NADH and L-lactate production were documented. The occurrence of such oscillations were found to depend mainly on the balance between the consumption of ATP associated with the phosphorylation of D-glucose, as catalyzed by both yeast and muscle hexokinase, and the net production of ATP resulting from the further catabolism of D-fructose 6-phosphate, as initiated by activation of phosphofructokinase. The oscillatory pattern was suppressed in the presence of D-fructose 2,6-bisphosphate. It is proposed that the quantitative information gathered in this study may set the scene for further studies in extracts of cells other than myocytes, e. g. hepatocytes and pancreatic islet cells, in which no oscillation of glycolysis was so far observed. 相似文献
6.
Branched chain alpha-ketoacid dehydrogenase and pyruvate dehydrogenase activity in isolated rat pancreatic islets 总被引:1,自引:0,他引:1
Branched-chain alpha-ketoacid dehydrogenase and pyruvate dehydrogenase in isolated rat pancreatic islets were shown to be regulated by a phosphorylation/dephosphorylation mechanism. Broad-specificity phosphoprotein phosphatase treatment stimulated and ATP addition inhibited their activities. The kinases responsible for inactivating these complexes were shown to be sensitive to inhibition by known inhibitors, alpha-chloroisocaproate and dichloroacetate. Total activity (nmol/min/islet / 37 degrees C) of branched-chain alpha-ketoacid dehydrogenase and pyruvate dehydrogenase was 0.86 and 5.09, with a % active form (activity before phosphatase treatment divided by activity after phosphatase treatment X 100) of 36% and 94%, respectively. Incubation of intact isolated islets with alpha-chloroisocaproate affected neither insulin release nor flux through branched-chain alpha-ketoacid dehydrogenase. 相似文献
7.
The coupling of metabolic to secretory events in pancreatic islets. The possible role of glutathione reductase 总被引:1,自引:0,他引:1
W J Malaisse S P Dufrane P C Mathias A R Carpinelli F Malaisse-Lagae P Garcia-Morales I Valverde A Sener 《Biochimica et biophysica acta》1985,844(2):256-264
The participation of glutathione reductase in the process of nutrient-stimulated insulin release was investigated in rat pancreatic islets exposed to 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). BCNU caused a time-and dose-related, irreversible inhibition of glutathione reductase activity. This coincided with a fall in both GSH/GSSG ratio and the thiol content of the islets. Pretreatment of the islets with BCNU inhibited the oxidation of glucose and its stimulant action upon both 45Ca net uptake and insulin release. Although BCNU (up to 0.5 mM) failed to affect the oxidation of L-leucine and L-glutamine, it also caused a dose-related inhibition of insulin release evoked by the combination of these two amino acids. The latter inhibition was apparently not fully accounted for by the modest to negligible effects of BCNU upon 45Ca uptake, 45Ca efflux, 86Rb efflux and cyclic AMP production. Since BCNU failed to inhibit insulin release evoked by the association of Ba2+ and theophylline, these results support the view that glutathione reductase participates in the coupling of metabolic to secretory events in the process of nutrient-stimulated insulin release. However, the precise modality of such a participation, for example the control of intracellular Ca2+ distribution, remains to be elucidated. 相似文献
8.
In rat parotid or pancreatic islet homogenates incubated at 7 degrees C, hexokinase displayed a greater affinity for but a lower maximal velocity with the alpha-anomer, as distinct from beta-anomer, of D-mannose. The anomeric specificity of mammalian hexokinase was similar in the case of D-mannose and D-glucose, but represented a mirror image of that of yeast hexokinase. 相似文献
9.
Impaired uptake of D-glucose by tumoral insulin-producing cells 总被引:1,自引:0,他引:1
At variance with the situation found in normal pancreatic islets, no equilibration of extracellular and intracellular D-glucose concentrations occurs in tumoral insulin-producing cells of the RINm5F line. This unexpected behaviour may account, in part at least, for the abnormal kinetics of glucose utilization in the tumoral cells and their poor secretory response to this hexose. 相似文献
10.
F Malaisse-Lagae M H Giroix A Sener W J Malaisse 《Biological chemistry Hoppe-Seyler》1986,367(5):411-416
The phosphorylation of alpha- and beta-D-glucose by either yeast hexokinase or beef heart hexokinase was measured at both 10 and 30 degrees C. At 30 degrees C, the anomeric specificity of yeast hexokinase represented a mirror image of that of bovine hexokinase, in terms of both maximal velocity and affinity. A decrease in temperature apparently accentuated the anomeric difference in both maximal velocity and affinity of bovine hexokinase. Such a difference consisted in a higher maximal velocity with beta- than alpha-D-glucose, but a greater affinity for the alpha- than beta-anomer. In yeast hexokinase, however, the decrease in temperature suppressed the anomeric difference in maximal velocity and inversed the anomeric difference in affinity. In the case of both enzymes, the fall in temperature decreased more the maximal velocity recorded with alpha-D-glucose than that measured with beta-D-glucose, and severely lowered the Km for alpha-D-glucose, whilst failing to affect significantly the Km for beta-D-glucose. These findings, which allow to reconcile prior apparently conflicting data, reveal that the anomeric behaviour of hexokinase is affected by the ambient temperature. Our data also support the view that hexokinase underwent a phylogenic evolution in terms of its anomeric specificity. 相似文献