A sensitive and specific assay for GTP and GDB in tissue extracts

Pogson, Christopher I.; Gurnah, Samantha V.; Smith, Stephen A.

doi:10.1016/0020-711x(79)90079-x

Cited by 8 publications

(6 citation statements)

References 29 publications

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“…As GTP has been demonstrated to attain concentrations of approx. 0.6 mm in the liver cell (Pogson et al, 1979), it is likely that this state will be physiologically relevant.…”

Section: Discussionmentioning

confidence: 99%

Guanosine 5′-triphosphate and guanosine 5′-[βγ-imido]triphosphate effect a collision coupling mechanism between the glucagon receptor and catalytic unit of adenylate cyclase

Houslay

Dipple

Elliott

1980

Biochemical Journal

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1. GTP, but not p[NH]ppG (guanosine 5'-[betagamma-imido]triphosphate), abolishes the sensitivity of glucagon-stimulated adenylate cyclase to the lipid-phase separations occurring in the outer half of the bilayer in liver plasma membranes from rat. 2. When either GTP or p[NH]ppG alone stimulate adenylate cyclase, the enzyme senses only those lipid-phase separations occurring in the inner half of the bilayer. 3. Trypsin treatment of intact hepatocytes has no effect on the basal, fluoride-, GTP- or p[NH]ppG-stimulated adenylate cyclase activity. However, (125)I-labelled-glucagon specific binding decays with a half-life matching that of the decay of glucagon-stimulated adenylate cyclase activity. 4. When GTP or p[NH]ppG are added to assays of glucagon-stimulated activity, the half-life of the trypsin-mediated decay of activity is substantially increased and the decay plots are no longer first-order. 5. Trypsin treatment of purified rat liver plasma membranes abolishes basal and all ligand-stimulated adenylate cyclase activity, and (125)I-labelled-glucagon specific binding. 6. Benzyl alcohol activates the GTP- and p[NH]ppG-stimulated activities in an identical fashion, whereas these activities are affected differently when glucagon is present in the assays. 7. We suggest that guanine nucleotides alter the mode of coupling between the receptor and catalytic unit. In the presence of glucagon and GTP, a complex of receptor, catalytic unit and nucleotide regulatory protein occurs as a transient intermediate, releasing a free unstable active catalytic unit. In the presence of p[NH]ppG and glucagon, the transient complex yields a relatively stable complex of the catalytic unit associated with a p[NH]ppG-bound nucleotide-regulatory protein.

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“…As GTP has been demonstrated to attain concentrations of approx. 0.6 mm in the liver cell (Pogson et al, 1979), it is likely that this state will be physiologically relevant.…”

Section: Discussionmentioning

confidence: 99%

Guanosine 5′-triphosphate and guanosine 5′-[βγ-imido]triphosphate effect a collision coupling mechanism between the glucagon receptor and catalytic unit of adenylate cyclase

Houslay

Dipple

Elliott

1980

Biochemical Journal

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“…The results were analysed graphically using Origin TM . 49 Commercial ADP (100 mg) was dissolved in TRIS buffer (6 ml, 0.1 M, pH 7.4), potassium chloride (6 ml, 50 mM), magnesium acetate (6 ml, 10 mM), and glucose (6 ml, 0.16 mM). Hexokinase (25 units) was added to this solution and the reaction was incubated for 30 min at rt.…”

Section: Determination Of K I Values For Inhibitorsmentioning

confidence: 99%

The synthesis of novel bisphosphonates as inhibitors of phosphoglycerate kinase (3-PGK)

Caplan

Pogson

Hayes

et al. 2000

J. Chem. Soc., Perkin Trans. 1

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“…One candidate for this role is phosphoenolpyruvate (PEP). 20). PEP also causes release of insulin from suspensions of isolated granules and plasma membranes 9,10.…”

mentioning

confidence: 99%

The role of phosphoenolpyruvate in insulin secretion: the effect of L-phenylalanine

et al. 1984

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A sensitive and specific assay for GTP and GDB in tissue extracts

Cited by 8 publications

References 29 publications

Guanosine 5′-triphosphate and guanosine 5′-[βγ-imido]triphosphate effect a collision coupling mechanism between the glucagon receptor and catalytic unit of adenylate cyclase

Guanosine 5′-triphosphate and guanosine 5′-[βγ-imido]triphosphate effect a collision coupling mechanism between the glucagon receptor and catalytic unit of adenylate cyclase

The synthesis of novel bisphosphonates as inhibitors of phosphoglycerate kinase (3-PGK)

The role of phosphoenolpyruvate in insulin secretion: the effect of L-phenylalanine

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