Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 4 de 4
Filter
Add more filters










Database
Language
Publication year range
1.
Biophys J ; 103(6): 1254-64, 2012 Sep 19.
Article in English | MEDLINE | ID: mdl-22995498

ABSTRACT

Conformational changes in the skeletal troponin complex (sTn) induced by rapidly increasing or decreasing the [Ca(2+)] were probed by 5-iodoacetamidofluorescein covalently bound to Cys-133 of skeletal troponin I (sTnI). Kinetics of conformational changes was determined for the isolated complex and after incorporating the complex into rabbit psoas myofibrils. Isolated and incorporated sTn exhibited biphasic Ca(2+)-activation kinetics. Whereas the fast phase (k(obs)∼1000 s(-1)) is only observed in this study, where kinetics were induced by Ca(2+), the slower phase resembles the monophasic kinetics of sTnI switching observed in another study (Brenner and Chalovich. 1999. Biophys. J. 77:2692-2708) that investigated the sTnI switching induced by releasing the feedback of force-generating cross-bridges on thin filament activation. Therefore, the slower conformational change likely reflects the sTnI switch that regulates force development. Modeling reveals that the fast conformational change can occur after the first Ca(2+) ion binds to skeletal troponin C (sTnC), whereas the slower change requires Ca(2+) binding to both regulatory sites of sTnC. Incorporating sTn into myofibrils increased the off-rate and lowered the Ca(2+) sensitivity of sTnI switching. Comparison of switch-off kinetics with myofibril force relaxation kinetics measured in a mechanical setup indicates that sTnI switching might limit the rate of fast skeletal muscle relaxation.


Subject(s)
Calcium/pharmacology , Myofibrils/drug effects , Myofibrils/metabolism , Troponin I/metabolism , Animals , Biomechanical Phenomena , Fluoresceins/metabolism , In Vitro Techniques , Kinetics , Muscle Relaxation/drug effects , Myofibrils/physiology , Psoas Muscles/drug effects , Rabbits , Sarcomeres/drug effects , Sarcomeres/metabolism
2.
Am J Physiol Cell Physiol ; 283(4): C1242-53, 2002 Oct.
Article in English | MEDLINE | ID: mdl-12225987

ABSTRACT

A countertransport of H(+) is coupled to Ca(2+) transport across the sarcoplasmic reticulum (SR) membrane. We propose that SR carbonic anhydrase (CA) accelerates the CO(2)-HCO reaction so that H(+) ions, which are exchanged for Ca(2+) ions, are produced or buffered in the SR at sufficient rates. Inhibition of this SR-CA is expected to reduce the rate of H(+) fluxes, which then will retard the kinetics of Ca(2+) transport. Fura 2 signals and isometric force were simultaneously recorded in fiber bundles of the soleus (SOL) and extensor digitorum longus (EDL) from rats in the absence and presence of the lipophilic CA inhibitors L-645151, chlorzolamide (CLZ), and ethoxzolamide (ETZ), as well as the hydrophilic inhibitor acetazolamide (ACTZ). Fura 2 and force signals were analyzed for time to peak (TTP), 50% decay time (t(50)), and their amplitudes. L-645151, CLZ, and ETZ significantly increased TTP of fura 2 by 10-25 ms in SOL and by 5-7 ms in EDL and TTP of force by 6-30 ms in both muscles. L-645151 and ETZ significantly prolonged t(50) of fura 2 and force by 20-55 and 40-160 ms, respectively, in SOL and EDL. L-645151, CLZ, and ETZ also increased peak force of single twitches and amplitudes of fura fluorescence ratio (R(340/380)) at an excitation wavelength of 340 to 380 nm. All effects of CA inhibitors on fura 2 and force signals could be reversed. ACTZ did not affect TTP, t(50), and amplitudes of fura 2 signals or force. L-645151, CLZ, and ETZ had no effects on myosin-, Ca(2+)-, and Na(+)-K(+)-ATPase activities, nor did they affect the amplitude and half-width of action potentials. We conclude that inhibition of SR-CA by impairing H(+) countertransport is responsible for deceleration of intracellular Ca(2+) transients and contraction times.


Subject(s)
Calcium/metabolism , Carbonic Anhydrase Inhibitors/pharmacology , Carbonic Anhydrases/metabolism , Muscle Fibers, Skeletal/metabolism , Muscle, Skeletal/enzymology , Acetazolamide/pharmacology , Action Potentials/drug effects , Action Potentials/physiology , Adenosine Triphosphatases/drug effects , Animals , Benzothiazoles , Chelating Agents/pharmacology , Dose-Response Relationship, Drug , Enzyme Activation/drug effects , Ethoxzolamide/pharmacology , Female , Fura-2/pharmacology , In Vitro Techniques , Ion Transport/drug effects , Ion Transport/physiology , Isometric Contraction/drug effects , Isometric Contraction/physiology , Muscle Contraction/drug effects , Muscle Contraction/physiology , Muscle Fibers, Skeletal/drug effects , Muscle, Skeletal/drug effects , Rats , Sarcoplasmic Reticulum/enzymology , Thiadiazoles/pharmacology , Thiazoles/pharmacology , Time Factors
3.
Pflugers Arch ; 443(5-6): 762-70, 2002 Mar.
Article in English | MEDLINE | ID: mdl-11889574

ABSTRACT

Muscle carbonic anhydrase (CA) was inhibited in fibre bundles of the extensor digitorum longus (EDL) and soleus (SOL) muscles from rats. Isometric single twitches were recorded in the absence or presence of the CA inhibitors. The highly membrane-permeable inhibitors L-645,151, chlorzolamide (CLZ) and ethoxzolamide (ETZ) prolonged significantly the values of time-to-peak (ttp) by 5-40 ms (10-40%) in both muscles and the values of the 75% decay time (t(75%)) by 30-400 ms (13-110%) in SOL and by 9-17 ms (15-30%) in EDL and increased peak force by 20--55% in SOL and EDL. The poorly membrane-permeable inhibitors benzolamide (BZ) and acetazolamide (ACTZ) had no effects on single twitches. In CO(2)-free solution, the effects of L-645,151 on ttp, t(75%) and peak force of SOL were reduced drastically. Removal of CO(2) prolonged ttp and t(75%). In skinned fibres, ETZ and CLZ did not increase force production. Intracellular pH (pH(i)) in SOL and EDL fibres was not affected by 30-60 min exposure to CLZ, ETZ or BZ. The results of L-645,151, CLZ and ETZ on ttp, t(75%) and peak force of twitches are consistent with our hypothesis on the role of the sarcoplasmic reticulum (SR) CA. The SR-CA may mediate sufficiently fast buffering and production of H(+) in the SR that is exchanged for Ca(2+) across the SR membrane. We propose that a H(+) buffering and delivery impaired by CA inhibition slows the kinetics of Ca(2+) release and reuptake and, as a result, slows twitch ttp and t(75%). Aspects of this hypothesis await further validation.


Subject(s)
Carbonic Anhydrase Inhibitors/pharmacology , Muscle Fibers, Skeletal/enzymology , Muscle Relaxation/physiology , Muscle, Skeletal/cytology , Thiazoles/pharmacology , Acetazolamide/pharmacology , Animals , Benzolamide/pharmacology , Benzothiazoles , Bicarbonates/pharmacology , Buffers , Carbon Dioxide/pharmacology , Carbonic Anhydrases/metabolism , Ethoxzolamide/pharmacology , Female , HEPES/pharmacology , Hydrogen-Ion Concentration , Muscle Contraction/drug effects , Muscle Contraction/physiology , Muscle Relaxation/drug effects , Rats , Rats, Inbred WF , Sarcoplasmic Reticulum/enzymology , Thiadiazoles/pharmacology
4.
News Physiol Sci ; 15: 269-274, 2000 Oct.
Article in English | MEDLINE | ID: mdl-11390925

ABSTRACT

Besides acting as an oxygen store during times of reduced blood oxygen supply, myoglobin can also facilitate intracellular oxygen transport by diffusion of oxymyoglobin along a PO(2) gradient. We reassess the importance of myoglobin-facilitated oxygen diffusion by applying new findings on the intracellular diffusivity of myoglobin in a model calculation.

SELECTION OF CITATIONS
SEARCH DETAIL
...