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Biomedical subjects

J Wolff

Publications and source records attributed to J Wolff.

At least 271 records · Page 15Linked to original sources
PubMed1974

Promotion of fluorescence upon binding of colchicine to tubulin.

Colchicine, which does not fluoresce in aqueous media and organic solvents, exhibits marked fluorescence on combination with brain tubulin, with a corrected excitation maximum at 362 nm, an emission maximum at 435 nm, and a quantum yield of about 0.03. From fluorescence measurements it was found that rat brain tubulin binds 0.83 moles of colchicine per dimer (molecular weight 110,000) with an association constant of 3.2 muM(-1) at pH 7.0 and 37 degrees . These results are in excellent agreement with those obtained with the binding of [(3)H]-colchicine. The enthalpy of binding is 10 kcal/mole, with an entropy change of 62 entropy units. The fluorescence can be ascribed to the tropolone moiety. However, the A ring of colchicine is also involved in binding. Denaturing agents abolish fluorescence, whereas podophyllotoxin, another antimitotic agent, decreases fluorescence competitively. Fluorescence is a convenient method for determining the binding of colchicine to tubulin that does not require the separation of free colchicine from bound colchicine and yields values for physical and biochemical parameters that are in excellent agreement with those obtained from the binding of [(3)H]colchicine.

Animals↗
PubMed1974

5'-Guanylylimidodiphosphate, a potent activator of adenylate cyclase systems in eukaryotic cells.

5'-Guanylylimidodiphosphate (Gpp(NH)-p) stimulates adenylate cyclase [ATP-pyrophosphate-lyase (cyclizing), EC 4.6.1.1] activity in plasma membranes isolated from frog and salmon erythrocytes, from rat adrenal, hepatic, and fat cells, and from bovine thyroid cells. The nucleotide acts cooperatively with the various hormones (glucagon, secretin, ACTH, thyrotropin, and catecholamines) that stimulate these adenylate cyclase systems with resultant activities that equal or exceed those obtained with hormone plus GTP or with fluoride ion. In the absence of hormones, Gpp(NH)p is a considerably more effective activator than GTP, and, under certain conditions of incubation, stimulates rat fat cell adenylate cyclase to levels of activity (about 20 nmoles of 3',5'-adenosine monophosphate mg protein per min) far higher than reported hitherto for any adenylate cyclase system examined. The nucleotide activates frog erythrocyte adenylate cyclase when the catecholamine receptor is blocked by the competitive antagonist, propranolol, and activates the enzyme from an adrenal tumor cell line which lacks functional ACTH receptors. In contrast, Gpp(NH)p does not stimulate adenylate cyclase in extracts from Escherichia coli B. Gpp(NH)p appears to be a useful probe for investigating the mechanism of hormone and nucleotide action on adenylate cyclase systems in eukaryotic cells.

Adenylyl Cyclases↗
PubMed1974

The contribution of subunits of thyroid stimulating hormone to the binding and biological activity of thyrotropin.

The binding of bovine TSH (thyroid stimulating hormone), LH (luteinizing hormone), and their subunits to the TSH receptor of beef thyroid membranes was compared to stimulation by these agents of adenylate cyclase [ATP pyrophosphate-lyase(cyelizing), EC 4.6.1.1] in the same membranes, glucose oxidation in dog thyroid slices, and the secretory process in mouse thyroids in vitro (colloid droplet formation) and in vivo (hormone release). The beta-subunits of TSH and LH can bind to the TSH receptor and can activate thyroid function in vitro. In contrast, the alpha-subunit of TSH binds negligibly to the TSH receptor and has very low potency for stimulation of thyroid function (except for colloid droplet formation). Neither binding nor the biological activity of the beta-subunits can be accounted for by TSH contamination, whereas this cannot be ruled out for alpha-TSH. LH binds to the TSH receptor even better than the beta-subunit of TSH but the increased binding does not result in a corresponding activation of thyroid function. Neither alpha- nor beta-TSH alone can induce more than 4-8% of the response to intact TSH in any of the investigated parameters. It is proposed that the beta-subunit has within its structure the primary determinants which are necessary to stimulate biological activity, whereas the alpha-subunit imposes conformational changes on the beta-subunit which in intact TSH promote binding and biological activity commensurately but in LH promote only binding.

Adenylyl Cyclases↗
PubMed1973

Stimulation of steroid secretion in adrenal tumor cells by choleragen.

Choleragen, the pure protein from cholera toxin, stimulates steroid secretion by Y-1 adrenal tumor cells in culture. The secreted steroids are the same as seen after addition of adrenocorticotropic hormone. Half-maximal stimulation occurs at 15 pM; stimulation is essentially irreversible by washing and partially reversible (for about 1 hr) by antibody, and there is a latent period of about 60 min before stimulation is seen. Stimulation of adenylate cyclase occurs at about 30-fold higher choleragen concentrations. Gangliosides inhibit choleragen stimulation when added before but not after the toxin. Lipopolysaccharides from Escherichia coli, Salmonella typhosa, and Serratia marcescens also stimulate steroid secretion, but are less potent than choleragen.

Adenylyl Cyclases↗
PubMed1972

Colchicine-binding protein and the secretion of thyroid hormone.

The role of microtubules in the thyrotropin- or adenosine 3',5' cyclic monophosphate (cyclic AMP)-stimulated accumulation of cytoplasmic colloid droplets and secretion of iodine from the mouse thyroid gland has been investigated by means of different classes of agents that affect the stability of microtubules. The onset of inhibition of secretion by colchicine, the uptake of colchicine-(3)H by thyroid lobes, and the binding of colchicine-(3)H to thyroidal soluble protein are shown to have similar time courses Colloid droplet accumulation is also inhibited and does not readily resume upon removal of colchicine from the medium. This appears to be due to the slow washout of the drug (t((1/2)) approximately hr). Thyroids contain a soluble colchicine-binding protein that resembles microtubule proteins of other tissues with respect to apparent K(m) for colchicine, pH optimum, and stability characteristics Colchicine analogues inhibit iodine secretion and colchicine binding in a parallel manner and as a function of their antimitotic potencies. Microtubule-stabilizing agents such as hexylene glycol and D(2)O also inhibit secretion. Thus, inhibition of thyroid secretion by antimitotic agents appears to be mediated by an effect on microtubules. The inhibitory locus of colchicine inhibition occurs after the generation of cyclic AMP, since stimulation of secretion by this nucleotide is blocked by colchicine, whereas thyroid-stimulating hormone-induced accumulation of cyclic AMP is not affected. Thus, the functioning microtubule appears to play a role in the induction of colloid endocytosis.

Animals↗
PubMed1972

The use of lithium in the treatment of thyrotoxicosis.

Since lithium has been shown to inhibit release of iodine from the thyroid, we have investigated its therapeutic potential in thyrotoxicosis. Eight detailed (131)I kinetic studies were performed on seven thyrotoxic women and data was analyzed using a computer program. Lithium at serum levels of about 1 mEq liter decreased the loss of (131)I from the thyroid, led to a fall in serum (131)I levels and diminished urinary (131)I excretion. Computer simulation of the lithium effect required, in every case, that lithium inhibit hormonal and nonhormonal thyroid iodine release. In five cases a second lithium effect was required for a satisfactory fit of the model soluton with observed data: namely, an inhibition of hormone disappearance from serum. NEITHER INHIBITION OF RELEASE NOR OF HORMONE DISAPPEARANCE SEEMED TO BE AFFECTED BY METHIMAZOLE (RELEASE: 52% decrease without methimazole, 60% with methimazole; hormone disappearance: approximately 60% decrease in both). When Li(+) was discontinued, recovery of the iodine release rate and hormone disappearance rate over the observed time span was variable, ranging from no recovery to rates that exceeded pre-Li(+) values. When Li(+) is used alone its effect on serum hormone levels is diminished due to continued accumulation of iodide by the thyroid. Thus, serum thyroxine-iodine levels fell 21-30% in 6-8 days in patients who did not receive methimazole and 15-67% in the methimazole-treated subjects. For prolonged therapy, therefore, a thiocarbamide drug must be used in conjunction with Li(+). The similarity of inhibition of iodine release from the thyroid produced by Li(+) and iodides is discussed.

Adolescent↗