Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UNIPROT:P01275 (glucagon)
 26,492 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The dietary and hormonal regulation of the level of glutathione-insulin tranhydrogenase in rat liver was investigated in these studies. In order to make valid comparisons, the assay of glutathione-insulin transhydrogenase was performed at near zero-order kinetics wherein enzyme rate was proportional to enzyme amount. Changing the protein content of the diet or administration of glucagon or cortisone did not significantly affect the specific activity of glutathione-insulin transhydrogenase in microsomes from rat liver. However, the Vmax and Km of this enzyme in the livers of adrenalectomized rats were increased three-and fourfold over these values in microsomes from normal liver. Administration of cortisone resulted in a return to the normal kinetic constants of microsomal GIT within 4 hr.
 ...
PMID:Environmental effects on glutathione-insulin transhydrogenase in rat liver. 96 93

Interactions of several proteins with glutathione-insulin transhydrogenase (GIT) have been investigated by determining their ability to inhibit degradation of 125I-labeled insulin catalyzed by GIT. The inhibition by every insulin analog (des-Asn-des-Ala-pork insulin, desoctapeptide-pork insulin, des-Ala-pork insulin, pork insulin, proinsulin, and guinea pig insulin) was competitive vs. competitive vs. insulin indicating that they function as alternate substrates. The insulin analogs with the least hormonal activity showed the highest potency as inhigitors of insulin degradation. Whereas native ribonuclease and lysozyme showed little or no inhibition, their scrambled forms (i.e. reduced and randomly reoxidized) showed competitive inhibition with a potency greater than that of insulin. These results suggest that the conformation of the substrate or inhibitor is probably the major factor in determining the specificity for (or binding to) the enzyme. Studies withother peptide hormones showed competitive inhibition with vasopressin and oxytocin and noncompetitive inhibition with glycagon. The inhibition with growth hormone could be either competitive or noncompetitive. The inhibition by glucagon and growth hormone (physiologic antagonists of insulin) could serve as a control mechanism to modulate the activity of enzyme. The following showed very little or no inhibition; the native and scrambled form of pepsinogen, trypsin inhibitor of beef pancreas and of lima bean, C-peptide of pork proinsulin, and heptapeptide (B23-B29) of insulin.
 ...
PMID:Interaction of insulin analogs, glucagon, growth hormone, vasopressin, oxytocin, and scrambled forms of ribonuclease and lysozyme with glytathione-insulin transhydrogenase (thiol: protein-disulfide oxidoreductase): dependence upon conformation. 117 Aug 77

Previously, we described primary cultures of rat liver cells in a serum-free multihormone defined medium (Am. J. Physiol. 243: E132-E151, 1982). This cell preparation exhibits marked insulin-dependent syntheses of protein, glycogen and lipids (hereafter collectively referred to as "bioactivity" for brevity). In the present studies, the role of different hormones in the expression of insulin's bioactivity was investigated. Hepatocytes from fasted rats, previously maintained in the multihormone glucagon-supplemented medium, were first cultured in the defined medium without the hormones and, subsequently, tested for bioactivity by replacing the missing hormones singly or in combination. Of all the hormones tested (testosterone, estradiol, thyroxine, glucocorticoid steroid and insulin), only insulin, dexamethasone and thyroxine when present individually in the culture medium, showed slight bioactivity (glycogenesis); however, insulin and dexamethasone, when present together, further increased each bioactivity, protein (1.3-fold), glycogen (8-fold) and lipid (1.6-fold) syntheses. The determination of apparent binding parameters of insulin specific receptors using Scatchard plots showed that insulin exposure caused a decrease in receptor concentration (Ro), dexamethasone exposure caused an increase in affinity (Kd), and, compared to insulin alone, treatment with insulin plus dexamethasone increased receptor concentration with no change in apparent affinity. Insulin induced a consistently small, but statistically significant, increase in the average specific activity of the protein-disulfide interchange enzyme, glutathione-insulin transhydrogenase (GIT), with equal distribution between its nonlatent ("readily available") and latent ("cryptic") forms. However, when dexamethasone was present along with insulin, the distribution of GIT was significantly greater in the latent form than in the nonlatent form. Examination by scanning and transmission microscopy showed clear differences, both in the cell surface and intracellular morphology, in the hepatocytes exposed to different hormonal milieu.
 ...
PMID:Role of insulin and dexamethasone in the expression of bioactivity in rat hepatocytes cultured in a serum-free defined medium. 389 65

Insulin degrading enzymes of rat liver cytosol, the so-called insulin and glucagon degrading proteinase (IGP, EC 3.4.23.5), and two forms of the insulin degrading thiol-protein-disulfide oxidoreductase/isomerase (glutathione-insulin transhydrogenase, TPO, EC 1.8.4.2/5.3.4.1) were separated from each other and partially purified on DEAE-Sephadex. The highly purified proteinase was obtained by polyacrylamide gel electrophoresis of the DEAE-Sephadex-purified enzyme fraction and was used to produce monospecific antibodies to the IGP in rabbits. Strong evidence is given that the insulin and glucagon degrading proteinase is an autonomous enzyme existing in addition to the TPO forms in the cytosol of the liver. Combined action of the proteinase and the TPO system on radioiodinated insulin under various conditions in vitro revealed an independent and non-sequential degradation of insulin by these two enzyme systems.
 ...
PMID:The insulin and glucagon degrading proteinase of rat liver. Separation of the proteinase from the thiol-proteindisulfide oxidoreductases. 637 96

Although previous studies from this and other laboratories have extensively characterized insulin degrading activity in animal tissues, little information has been available on insulin responsive human tissues. The present study describes the insulin degrading activity in skeletal muscle from normal human subjects. Fractionation of a sucrose homogenate of skeletal muscle demonstrated that 97% of the total neutral insulin degrading activity was in the 100 000 x g supernatant with no detectable glutathione-insulin transhydrogenase activity. The 100000 x g pellet contained 85% of the total acid protease activity and all the glutathione-insulin transhydrogenase activity. The soluble insulin degrading activity was purified 1400-fold by ammonium sulfate fractionation, molecular exclusion, ion-exchange and affinity chromatography. Enzymatic activity was determined by measuring an increase in trichloroacetic acid-soluble products of the 125I-labeled hormone substrates. The purified enzyme showed marked proteolytic specificity for insulin with a Km of 1.63 X 10(-7)M (+/-0.32) and was competitively inhibited by proinsulin and glucagon with Ki values of 2.1 X 10(-6)M and 4.0 X 10(-6)M, respectively. This insulin protease exhibited a pH optimum between 7 and 8, a molecular weight of 120000 and was capable of degrading glucagon. Inhibition studies demonstrated that a sulfhydryl group is essential for activity. Molecular exclusion chromatography of [125I]insulin degraded products revealed a time-dependent increase in degradation products with molecular weights intermediate between intact insulin and iodotyrosine. These studies demonstrate that the major enzymatic system responsible for insulin degrading activity is a soluble cysteine protease capable of rapidly metabolizing insulin under physiologic conditions.
 ...
PMID:Insulin degradation by human skeletal muscle. 675 60

Native insulin inhibits the binding and degradation of (125)I-labelled insulin in parallel. Half-maximal inhibition of degradation occurs with 10nm-insulin, a hormone concentration sufficient to saturate the insulin receptor. The proportion of bound hormone that is degraded increases as the insulin concentration is increased, suggesting that low-affinity uptake is functionally related to degradation. Since only a small fraction (approx. 10%) of the overall degradation occurs at the plasma membrane, or in the extracellular medium, translocation of bound hormone into the cell is the predominant mechanism mediating the degradation of insulin. In the presence of 0.6nm-insulin, a concentration at which most cell-associated hormone is receptor-bound, chloroquine increases the amount of (125)I-labelled insulin retained by hepatocytes. However, chloroquine increases the retention of degradation products of insulin in incubations containing sufficient hormone (6nm) to saturate the receptor and permit occupancy of low-affinity sites. Glucagon does not compete for the interaction of (125)I-labelled insulin (1nm) with the insulin receptor. In contrast, 20mum-glucagon inhibits 75% of the uptake of insulin (0.1mum) by low-affinity sites. A fraction of the cell-bound radioactivity is not intact insulin throughout a 90min association reaction at 37 degrees C. During dissociation, fragments of (125)I-labelled insulin are released to the medium more rapidly than is intact hormone. The production and transient retention of degradation products of the hormone complicates the characterization of the insulin receptor by equilibrium or kinetic methods of assay. It is proposed that insulin degradation occurs by receptor- and non-receptor-mediated pathways. The latter may be related to the action of glutathione-insulin transhydrogenase, with which both insulin and glucagon interact.
 ...
PMID:Receptor- and non-receptor-mediated uptake and degradation of insulin by hepatocytes. 676 Aug 55