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M Tessari

Publications and source records attributed to M Tessari.

26 records · Page 2Linked to original sources
PubMed1992

Conformation of uteroglobin fragments.

The conformation of three fragments of uteroglobin in aqueous solution and in the presence of SDS micelles is described. Two of these fragments correspond to helix II and helix III of uteroglobin, the crystal structure of which is made of four helices. The third peptide comprises helices II and III, with the connecting beta-turn. While helix II does not interact strongly with the micelles, helix III adopts a rather clear alpha-helix in this system. The elongation of helix III with the addition of helix II at the N-terminus somewhat stabilizes the ordered structure. It is possible that the beta-turn found in the crystal is also present in solution.

Amino Acid Sequence↗
PubMed1991

Na(+)-Ca2+ exchange activity in synaptic plasma membranes derived from the electric organ of Torpedo ocellata.

Synaptic plasma membranes obtained by hypo-osmotic treatment of purified Torpedo ocellata synaptosomes, contain an electrogenic Na(+)-Ca2+ exchange system. The dependence of the initial reaction rate on [Ca2+] reveals a single binding site for Ca2+ with an average apparent Km of 13.66 (S.D. = 12.07) microM [Ca2+] and maximal reaction velocity of Vmax = 11.33 (S.D. = 5.93) nmol/mg protein per s. The dependence of the initial rate of the Na+ gradient dependent Ca2+ influx on the internal [Na+] exhibits a sigmoidal curve which reaches half-maximal reaction rate at 170.8 (S.D. = 19.9) mM [Na+]. Addition of ATP gamma S does not change the K0.5 to Na+. The average Hill coefficient is 3.09 (S.D. = 0.86) indicating that 3-4 Na+ ions are exchanged for each Ca2+. Na+ gradient dependent Ca2+ uptake in Torpedo SPMs takes place also in the absence of K+ suggesting that K+ co-transport is not obligatory. The temperature dependence of the initial and steady-state rates of Na+ gradient dependent Ca2+ influx reveal that maximal reaction velocities of the Torpedo exchanger are attained between 15 and 20 degrees C. The energy of activation between 0 and 20 degrees C is 20,826 cal/mol. In comparison, rat brain synaptic plasma membrane Na(+)-Ca2+ exchanger reaches maximal reaction rates between 30 and 40 degrees C. Reconstitution of Torpedo or rat brain Na(+)-Ca2+ exchangers into a membrane composed of either Torpedo or brain phospholipids, does not alter the temperature dependence of the native Torpedo or rat brain Na(+)-Ca2+ exchangers; inspite of considerable differences in the composition of the fatty acyl chains that are esterified to brain and Torpedo phospholipid head groups and differences in membrane fluidity that were detected. An ATP-dependent Ca2+ pump, which is insensitive to FCCP, is also present in the same synaptic membrane.

Animals↗
PubMed1991

Molecular and mechanistic heterogeneity of the Na(+)-Ca2+ exchanger.

1. Studying the effect of K+ on Na(+)-Ca2+ exchange in rat brain SPMs revealed that a consistent stimulation was obtained. This stimulation persisted also when FCCP was included in the K(+)-containing reaction mixture to minimize the effect of membrane potential on the electrogenic process. 2. Using Rb+ as a K+ analogue revealed that it was cotransported with Ca2+ in a Na+ gradient-dependent manner. The ratio between the amount of Ca2+/Rb+ transported in rat brain SPMs in a Na+ gradient-dependent manner suggests that not all the Na(+)-Ca2+ exchangers in that preparation cotransport Rb+ (K+) with Ca2+. This is supported also by the finding that Na+ gradient-dependent Ca2+ influx can proceed in rat brain SPMs in the complete absence of K+ although to a lesser extent. 3. Protein purification studies and immunological characterization indicate that a 70-kDa protein is consistently detected in rat brain SPMs. Immunological characterization of the proteins expressed in the 14-18 S mRNA-injected Xenopus oocyte in conjunction with Na+ gradient dependent Ca2+ uptake activity or in the same mRNA-fortified reticulocyte lysate suggest that proteins of about 70 kDa are specifically synthesized. 4. Torpedo electric organ Na(+)-Ca2+ exchanger differs at least in two respects from the rat brain Na(+)-Ca2+ exchanger: It has a low affinity to Na+ (K0.5 = 170 mM), and it reaches maximal activity between 15-20 degrees C. Reconstitution studies suggest that the temperature difference might reflect a difference in the proteins themselves rather then a difference in membrane fluidity due to a difference in the membrane lipid composition.

Animals↗
PubMed1989

Inhibition of Na+/Ca2+ exchange by amiloride acting from opposite sides of cardiac sarcolemma.

Amiloride inhibited the Na+Ca2+ exchange activity of cardiac sarcolemmal vesicles with similar affinities at the cis and trans sides of the membrane, estimated apparent Ki on both sides of the sarcolemma being similar. The extent of amiloride inhibition on Na+/Ca2+ exchange activity was decreased by alkaline pH only when the drug was acting from the external side of the vesicle sarcolemma, whereas when vesicles were preincubated with the drug at different pH values, amiloride appeared to act as a weak permeant base, being a more effective inhibitor at alkaline pH values. In fact, a rise in the pH of the preincubation medium may favour the entry and consequently the effect of the drug on the exchanger. The pH dependence of the inhibition of Na+/Ca2+ exchange activity by either extravesicular or intravesicular amiloride was consistent with the hypothesis that in both cases the protonated drug was the active form. Evidence is presented that the pattern of interaction of amiloride on the Na+/Ca2+ exchange system strictly depended on the sidedness of drug action. In fact, while Na+ protected against inhibition by amiloride when it was acting on the same side of the vesicle membrane as the drug, it synergically interacted with amiloride to inhibit exchange activity when it was acting on the opposite side of the sarcolemma as the drug. Furthermore, only extravesicular amiloride removed the stimulation of Na+/Ca2+ exchange activity in Ca2+-treated vesicles.

Amiloride↗
PubMed1987

Effects of N-chlorobenzyl analogues of amiloride on myocardial contractility, Na-Ca-exchange carrier and other cardiac enzymatic activities.

1. In electrically driven guinea pig left atria, micromolar concentrations (2 mumol/l to 80 mumol/l) of N-chlorobenzyl derivatives of amiloride (o-chlorobenzamil and 3',4'-dichlorobenzamil) produced quantitatively similar positive inotropic effects. Contracture developed with 3',4'-dichlorobenzamil. Endogenously released catecholamines contributed 30% to the positive inotropic effect of o-chlorobenzamil but did not contribute at all to the effect of 3',4'-dichlorobenzamil. When tested in the presence of the inhibitor of phosphodiesterase isobutylmethylxanthine, o-chlorobenzamil antagonized its positive inotropic effect, whereas 3',4'-dichlorobenzamil potentiated it. o-Chlorobenzamil also antagonized the positive inotropic effect of ouabain in that it shifted its concentration-effect curve to the right. Moreover, o-chlorobenzamil prevented the appearance of ouabain toxicity in terms of a rise in the resting force. 2. Also, in electrically driven guinea pig papillary muscle, micromolar concentrations (5 mumol/l to 30 mumol/l) of both N-chlorobenzyl derivatives of amiloride produced a positive inotropic effect. This effect was more marked with 3',4'-dichlorobenzamil than with o-chlorobenzamil and was associated for both compounds with lengthening of relaxation time. 3. o-Chlorobenzamil and 3',4'-dichlorobenzamil influenced, though not to the same extent, several systems involved in the onset and in the control of cardiac contractility. 3',4'-Dichlorobenzamil inhibited with the same potency Na-K-ATPase, sarcotubular Ca-ATPase, Na-Ca-exchange carrier, cAMP-dependent phosphodiesterase isolated from bovine heart and oxidative phosphorylation of mitochondria isolated from rat liver. Low micromolar concentrations of o-chlorobenzamil mainly inhibited Na-Ca-exchange carrier and cAMP-dependent phosphodiesterase.(ABSTRACT TRUNCATED AT 250 WORDS)

Amiloride↗
PubMedDate not supplied

Neural substrate of nicotine addiction as defined by functional brain maps of gene expression.

The distributed neural networks involved in the intravenous self-administration of nicotine and cocaine, and in a model of relapse of nicotine-taking after abstinence, were compared in Wistar rats. Post-mortem brain maps of c-fos-related antigens expression showed specific activation in prefrontal cortex, anterior cingulate and nucleus accumbens for both drugs, but of the anterior cingulate cortex only during relapse, suggesting that a subset of the neural network involved in drug self-administration is activated during relapse.

Animals↗