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

Recombinant human glucagon was successfully produced with a high level of expression in Escherichia coli as a fusion protein with human interferon gamma. The synthetic gene was designed to release glucagon, which does not contain glutamic acid residues, from fusion protein with the Staphylococcus aureus strain V8 protease that specifically cleaves the peptide bond on the carboxyl side of the glutamic acid residue. The resulting glucagon was purified to homogeneity by a combination of C18 reverse-phase HPLC and ion-exchange HPLC. The yield of intact glucagon obtained from 11 of culture was approximately 12 mg. The structure of recombinant human glucagon was confirmed by HPLC and amino acid composition/sequence analyses.
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PMID:Production of recombinant human glucagon in the form of a fusion protein in Escherichia coli; recovery of glucagon by sequence-specific digestion. 136 1

The thermodynamic behaviour of three peptides, bombesin, beta-endorphin and glucagon, was studied under reversed-phase high-performance liquid chromatographic conditions. Experimental data related to the interactive surface contact area (S values) and solute affinity (log k0) were derived over a range of temperatures between 5 and 85 degrees C. These experimental conditions allowed changes in the secondary structure of the solute to be monitored. The influence of the nature of the stationary phase ligand on the relative conformational stability of the three peptides was analysed by acquiring data with n-octadecyl silica (C18) and n-butyl silica (C4) sorbents. Values for the relative changes in entropy and enthalpy associated with the interactive process were also determined. The results provide further insight into the factors involved with the stabilization of secondary structure and the mechanism of the interaction of peptides with hydrophobic surfaces.
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PMID:High-performance liquid chromatography of amino acids, peptides and proteins. CXV. Thermodynamic behaviour of peptides in reversed-phase chromatography. 163 93

After preparative high-performance liquid chromatography of mouse islet culture medium, concentrated on disposable C18 cartridges (Sep-Pak), an unexpected insulin immunoreactive peak eluting earlier than mouse insulin I and II was detected. Molecular mass determination by mass spectrometry supported its suspected identity as methionine sulphoxide insulin II. We have examined the formation of Met-O derivatives of insulin II, glucagon and pancreatic polypeptide during sample preparation (Sep-Pak and Speed-Vac concentrating). The oxidation of methionine residues was found to depend very much on the buffer, the organic modifier and the procedure. In particular the use of methanol-trifluoroacetic acid resulted in extensive oxidation. The oxidation could be minimized by adding 2 mM dithiothreitol to the buffer and by degassing and/or nitrogen-bubbling of the buffer. Minimal formation of Met-O derivatives is important for the quantitation of methionine-containing polypeptides.
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PMID:High-performance liquid chromatography of rat and mouse islet polypeptides: potential risk of oxidation of methionine residues during sample preparation. 227 17

Male Sprague-Dawley rats were fed diets varying in fatty acid composition for 24 d. Liver plasma membranes were isolated, and the effect of diet on phospholipid fatty acyl tail composition and glucagon-stimulated adenylate cyclase activity was measured. Dietary linolenic acid influenced membrane phospholipid fatty acid composition and altered the effect of different dietary levels of linoleic acid on membrane composition. At low dietary intakes of linolenic acid, membrane fatty acids derived from linolenic acid increased as dietary intake of C18:2(9,12) increased. At high dietary linolenic acid levels membrane content of fatty acids derived from linolenic acid decreased as dietary intake of linoleic acid increased. Glucagon-stimulated adenylate cyclase activity decreased at high levels of both dietary linoleic acid and linolenic acid. These observations suggest that dietary balance between linoleic and linolenic acids has a role in plasma membrane composition and may control glucagon-stimulated adenylate cyclase activity.
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PMID:Diets varying in linoleic and linolenic acid content alter liver plasma membrane lipid composition and glucagon-stimulated adenylate cyclase activity. 287 99

In an effort to find analogs of glucagon that would bind to the glucagon receptor of the rat liver membrane but would not activate membrane-bound adenyl cyclase, several hybrid molecules were synthesized which contained sequences from both glucagon and secretin. [Asp3, Glu9]Glucagon and [Asp3, Glu9, Arg12]glucagon were inactive in the adenyl cyclase assay even at high concentrations but retained some binding affinity for the receptor. They were able to displace 125I-glucagon completely from its receptor and could completely inhibit the activation of adenyl cyclase by natural or synthetic glucagon. The inhibition index [I/A]50 was approximately 110 for both analogs. [Asp3]Glucagon, [Glu3]glucagon and [Asp3, Lys17, 18, Glu21]glucagon were weak partial agonists, while [Asp3, Glu21]glucagon was inactive and a poor inhibitor. The peptides were synthesized by solid-phase methods and purified to homogeneity by reverse-phase high-performance liquid chromatography on C18 silica columns. These are the first fully synthetic competitive glucagon antagonists to be reported.
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PMID:Glucagon antagonists. Synthesis and inhibitory properties of Asp3-containing glucagon analogs. 303 23

Several glucagon analogs were synthesized in an effort to find derivatives that would bind with high affinity to the glucagon receptor of rat liver membranes but would not activate membrane-bound adenylate cyclase and, therefore, would serve as antagonists of the hormone. Measurements on a series of glucagon/secretin hybrids indicated that replacement of Asp9 in glucagon by Glu9, found in secretin, was the important sequence difference in the N terminus of the two hormones. Further deletion of His1 and introduction of a C-terminal amide resulted in des-His1-[Glu9]glucagon amide, which had a 40% binding affinity relative to that of native glucagon but caused no detectable adenylate cyclase activation in the rat liver membrane. This antagonist completely inhibited the effect of a concentration of glucagon that alone gave a full agonist response. It had an inhibition index of 12. The pA2 was 7.2. An attempt was made to relate conformation with receptor binding. The peptides were synthesized by solid-phase methods and purified to homogeneity by reverse-phase high-performance liquid chromatography on C18-silica columns.
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PMID:Synthetic peptide antagonists of glucagon. 303 68

Rat and human hepatocyte cultures were incubated with 5 common plasma longchain fatty acids (C16-C18). Rates of fatty acid uptake were similar in rat and human hepatocytes and were of the order: 16:1 greater than 16:0; 18:2 greater than 18:1 greater than 18:0. Rates of ketogenesis were lower in human compared to rat hepatocytes. In rat hepatocytes glucagon stimulated ketogenesis only in the presence of exogenous carnitine and rates of ketogenesis were higher from unsaturated compared to corresponding saturated fatty acids. Glucagon decreased triacylglycerol secretion irrespective of the fatty acid substrate and it increased intracellular triacylglycerol accumulation. The latter effect of glucagon was more marked in the absence of carnitine supplementation.
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PMID:Fatty acid uptake and metabolism to ketone bodies and triacyglycerol in rat and human hepatocyte cultures is dependent on chain-length and degree of saturation. Effects of carnitine and glucagon. 340 19

2-Nitro-4-azidophenylsulphenyl-glucagon, a specific photoaffinity label for glucagon receptors, was synthesized and radioactively labelled with 125I. The radio-labelled peptide was purified from the reaction mixture by high-performance liquid chromatography in one step by isocratic elution from a C18 column with 20.4% n-propanol in 10 mM phosphate buffer (pH 2.5) as eluent. This glucagon derivative can be used to attach a label specifically to the glucagon receptor. The binding ability of the photoaffinity derivative was tested on isolated intact rat hepatocytes. Compared with a Kd of 3 nM for unmodified monoiodinated glucagon, the Kd value of the photoaffinity labelled monoiodinated glucagon tracer was 7 nM.
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PMID:Purification of radioiodinated photoactivable glucagon by isocratic high-performance liquid chromatography. 404 35

Platelet function, estimated from plasma beta-thromboglobulin (beta-TG, ng/ml), is frequently altered in insulin-dependent diabetics (IDDs). As several factors may affect beta-TG, we studied respectively in 15 IDDs, the roles played by: (i) diabetic control evaluated from glycosylated haemoglobin (HbA1); (ii) plasma C-peptide and pancreatic glucagon; (iii) plasma lipids and the relative percentages of fatty acids in total plasma lipids. Plasma beta-TG did not correlate significantly with the first 3 parameters. However, beta-TG was correlated: (i) positively with plasma triglycerides (P less than 0.01), cholesterol (P less than 0.02), phospholipids (P less than 0.05) and total plasma lipids (P less than 0.01) and the percentage of oleic acid (C18 : 1 omega 9) in plasma lipids (P less than 0.01); (ii) negatively with the percentage of linoleic acid (C18 : 2 omega 6) in plasma lipids (P less than 0.02). No correlation was found between beta-TG and the percentages of the other saturated (C16 : 0, C18 : 0), monounsaturated (C16 : 1 omega 7) and polyunsaturated fatty acids (C18 : 3 omega 6, C20 : 3 omega 6 and C20 : 4 omega 6). The present results indicate that beta-TG in IDDs can be markedly improved by all dietary and therapeutic measures which lower plasma lipids and increase the percentage of the linoleic acid in the body.
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PMID:Plasma lipid fatty acids and platelet function in insulin-dependent diabetic patients. 623 Feb 69

The synthesis of [125I-Tyr10]monoiodoglucagon from glucagon and carrier-free 125I using 1,3,4,6-tetrachloro-3-6-diphenylglycouril (Iodogen) and its separation in pure form by reverse phase high pressure liquid chromatography (HPLC) over C18-muBondapak columns using two consecutive linear gradients between solvent A [40:60 mixture of methanol and 10 mM H3PO4 in H2O (pH adjusted to 3.0 with triethylamine)] and solvent B (50:50 mixture of acetonitrile and 0.1 M Tris-HCl, pH 9.0) is reported. The newly synthesized [125I]monoiodoglucagon is shown to activate adenylyl cyclase in liver membranes with an EC50 between 5- and 8-fold lower than that of native glucagon. Further, it binds specifically to sites on liver plasma membranes that have the characteristics of glucagon receptors in terms of guanine nucleotide sensitivity and rates of reaction. It is suggested that [125I-Tyr10]monoiodoglucagon is a suitable probe for studying structural and functional properties of glucagon receptors.
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PMID:Monoiodoglucagon: synthesis, purification by high pressure liquid chromatography, and characteristics as a receptor probe. 630 49


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