Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.6.1.3 (ATPase)
 65,361 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The 38-kDa chymotryptic fragment of caldesmon, which possesses the actin/calmodulin binding domain, was purified and utilized to study the mechanism for the inhibition of acto-myosin ATPase by caldesmon. The intact caldesmon inhibited the acto-HMM ATPase although it caused an increase in the binding of HMM to actin, presumably due to the interaction between the S-2 region of HMM and the caldesmon located on the actin filament. The 38-kDa fragment, which lacks the S-2 binding domain, inhibited both the acto-HMM ATPase and the HMM binding to actin. The ATPase and the HMM binding to actin decreased in parallel on increasing the 38-kDa fragment bound to actin. In the presence of tropomyosin, the ATPase activity fell more rapidly than did the HMM binding to actin. Binding of intact caldesmon or 38-kDa fragment to actin inhibited the cooperative turning-on of tropomyosin-actin by NEM.S-1, which forms rigor complexes in the presence of ATP. The absence of cooperative turning-on of the acto-HMM ATPase by rigor complexes in the presence of 38-kDa fragment was associated with an inhibition of the binding of HMM to tropomyosin-actin. Addition of NEM.S-1 to tropomyosin-actin-caldesmon caused a gradual decrease in the caldesmon-induced binding of HMM to actin. The calmodulin restored the caldesmon-induced binding of HMM to tropomyosin-actin, but it had only a slight effect on the acto-HMM ATPase. These data suggest that the cooperative turning-on of the smooth muscle tropomyosin-actin by rigor bonds is modulated by the interaction of caldesmon, tropomyosin, and calmodulin on the thin filament.
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PMID:Caldesmon inhibits the cooperative turning-on of the smooth muscle heavy meromyosin by tropomyosin-actin. 253 47

We have developed a new method to prepare single-headed heavy meromyosin with high purity and a high yield. To examine whether the two heads on the same myosin molecule work cooperatively or not, it is important to prepare pure single-headed heavy meromyosin. Myosin was extracted from myofibrils treated with a solution containing CyDTA, a strong divalent cation chelator. CyDTA treatment was essential to the production of sHMM. Then such myosin was digested with chymotrypsin in the presence of divalent cations at high ionic strength. Crude sHMM was separated from double-headed HMM by affinity chromatography using an ADP-column. Contaminating S1 was removed by gel filtration. Heavy chain of sHMM obtained by the present method had no nick. Purified sHMM showed normal EDTA-ATPase and Ca-ATPase. It interacted with thin filament and its ATPase was activated by actin normally.
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PMID:New method to prepare single-headed heavy meromyosin with high purity and a high yield. 253 47

The in vitro effects of piperine on three bioenergetic reactions namely, oxidative phosphorylation, ATPase activity and calcium transport by isolated rat liver mitochondria have been investigated. Piperine was found to inhibit state 3 and DNP-stimulated respiration by mitochondria respiring with glutamate plus malate or succinate as substrates. The I50 values of piperine on oxidative phosphorylation in the presence of glutamate plus malate and succinate were 22 and 12 micrograms/mg mitochondrial protein respectively. With HTM preparations, the oxidation of added NADH and succinate was depressed by piperine while ascorbate plus TMPD oxidation was slightly affected. Piperine did not inhibit the mitochondrial ATPase activity induced by DNP, but by itself exerted stimulating activity on this enzyme. Piperine was also found to diminish calcium uptake and to facilitate the release of accumulated calcium by the mitochondria incubated with succinate or ATP. These results suggest that piperine inhibits mitochondrial oxidative phosphorylation at the level of respiratory chain, and the inhibitory site(s) is in the segment(s) ahead of cytochrome C. The mechanism of the piperine-induced ATPase activity is not known; but the effect of piperine on calcium transport is likely to be consequential to the effects of this compound on the mitochondrial respiratory chain and ATPase activity.
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PMID:Effects of piperine on bioenergetic functions of isolated rat liver mitochondria. 296 41

Myosin has two heads which can bind with F-actin and react with ATP. The skeletal muscle myosin forms each 1 mol of the myosin-phosphate-ADP complex (M-P-ADP) and the myosin-ATP complex (M-ATP). The actomyosin ATPase reaction which is coupled with muscle contraction is catalyzed only by the head which forms M-P-ADP. However, the function of M-ATP forming head in muscle contraction has not been elucidated. We studied the binding of S-1 and HMM with F-actin and the dissociation of acto-S-1 or acto-HMM by ATP or AMPPNP using the change in light-scattering and fluorescence of pyrene bound to F-actin. S-1 and HMM bound with actin at 1:1 and 1:2 molar ratio, respectively. Acto-S-1 dissociated by one mole of ATP per mole of S-1 but acto-HMM dissociated by 1 mol ATP per mol of HMM (0.5 mol/mol head). Acto-HMM dissociates by AMPPNP (or ADP) via a ternally complex. Acto-HMM bound two mole of AMPPNP, but acto-HMM dissociated by a function of (AMPPNP) but not (AMPPNP)2. These results suggested that the affinity of HMM with F-actin decreased by the binding of one mole of AMPPNP. The result presented here showed that binding of M-ATP forming head with F-actin is controlled by the ATPase reaction of the M-P-ADP forming head. It is suggested that during muscle contraction two heads react cooperatively with thin filament.
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PMID:The function of two heads of myosin in muscle contraction. 297 Feb 8

Activation of aorta thiophosphorylated heavy meromyosin (HMM[SP]) Mg2+-ATPase activity by aorta actin and the fraction of HMM[SP]-substrate intermediate complexes bound to actin were measured simultaneously. At 25 degrees C the Km for ATPase activation and the dissociation constant for the binding reaction were similar, irrespective of the presence or absence of tropomyosin. Aorta caldesmon (0.1 mol/mol actin) inhibited ATPase activation by 80-90% but did not alter the binding of HMM[SP]-product intermediates to actin. It is concluded that caldesmon inhibits by slowing the rate-limiting release of products from the actin-HMM[SP].ADP.Pi complex.
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PMID:Aorta caldesmon inhibits actin activation of thiophosphorylated heavy meromyosin Mg2+-ATPase activity by slowing the rate of product release. 297 72

In striated muscle myosin, a proteolysis site at the 25-50 kDa junction, susceptible in the filament and efficiently protected by nucleotides, is similarly protected when myosin is monomeric. Kinetic studies at low ionic strength show a close relationship between LC2 cleavage or degradation rate and cleavage of the 25-50 kDa heavy chain site. The myosin-[(T)-LC2'] species forms normal reconstituted filaments but its 25-50 kDa site susceptibility is closer to that of monomeric myosin, thus becoming practically ionic strength-independent. In this species the absence of the LC2 N-terminal segment induces a significantly greater susceptibility of the papain-sensitive site in LC1. In an LC2-depleted myosin the 25-50 kDa site susceptibility also becomes ionic strength-independent, however, the cleavage rates are then closer to that of filaments. Susceptibility in HMM and S1 is also much less dependent on ionic strength with rates intermediary between those of filament and monomer. These observations show that the maximum susceptibility to papain of the 25-50 kDa site requires both the integrity of the LC2 light chain and the filament structure and furthermore provide evidence that: (i) the LC2 N-terminus interacts specifically with some part of the filament; (ii) this interaction induces a specific transconformation in a region close to the ATPase active site; (iii) there is an interrelationship between LC1 and LC2 light chain N-terminal extremities, at least in the filament structure.
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PMID:Proteolysis rates of a myosin heavy chain site with papain. Evidence for a combined LC2-filament-mediated mechanism. 330 15

Crude extracts obtained by water extraction from the muscle of an ascidian (Halocynthia rotezi) are shown to possess an inhibitory activity on ATPase [(Na, K) ATPase, TF1-ATPase, HMM-ATPase]. Gel filtration of the extract with Sephadex G-10 separated the activity into two peaks, and the latter peak (inhibitor II) was further purified by HPLC. This inhibitor was non-competitive for ATP and associated firmly with the (Na, K) ATPase.
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PMID:[ATPase inhibitory activities found in the soluble fraction of an ascidian (Halocynthia rotezi) muscle]. 357 8

The effects of nucleotides and Ca2+ on the intrinsic tryptophan fluorescence of molluscan myosin and its proteolytic fragments were studied. By using these proteins from the scallop, Pecten maximus, the existence of two distinct tryptophan-containing domains was established, which respond independently to ATP and Ca2+-specific binding. The latter is located in the 'neck' region of the myosin, which constitutes the regulatory domain. Subfragment 1, lacking the regulatory domain, responded only to ATP binding. On the other hand a tryptic fragment comprising the regulatory domain responded only to Ca2+ binding. Subfragment 1, containing the regulatory domain, responded to both ATP and Ca2+, but its ATPase activity was Ca2+-insensitive. By contrast, the ATPase activity of HMM was Ca2+-sensitive. Increasing the ionic strength had a detrimental effect on Ca2+-sensitivity, and fluorescence studies on solubilized myosin were therefore of limited value. Myosin and its fragments from other molluscan species which were investigated produced similar changes to those of Pectan maximus.
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PMID:Fluorescence studies on the nucleotide- and Ca2+-binding domains of molluscan myosin. 390 36

Two different HMM species of gizzard myosin were prepared under conditions such that the phosphorylation of light chain was fully maintained. They were different in the N-terminal structure of the heavy chain but not in the light chain composition. A significant decrease in the Mg2+-ATPase activity was observed in one class of HMM which was proteolytically cleaved intramolecularly at site 1, 5 K daltons from the masked N terminus. Another class of HMM without the cleavage at site 1 showed ATPase activity similar to that of myosin. The decrease in ATPase activity was not caused by denaturation since similar amounts of initial burst of Pi liberation were observed with both HMMs and myosin. Kinetic and substructure analyses of HMM revealed that the activity change depended solely on the cleavage at site 1. The N-terminal region of gizzard myosin heavy chain may thus have an important role in maintaining the active site structure.
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PMID:N-terminal region of gizzard myosin heavy chain is critical for the ATPase activity. 611 61

The obliquely striated body wall muscle of the earthworm Lumbricus terrestris L. possesses a dual actin-linked and myosin-linked regulatory system. Tropomyosin from this muscle has now been purified and its functional properties compared to tropomyosin from vertebrate skeletal muscle. Earthworm tropomyosin has a molecular weight of about 70 000 and is composed of two polypeptide chains of molecular weight of 34 000 and 37 000. Structural and functional similarities to skeletal muscle tropomyosin were demonstrated with respect to the formation and periodicity of paracrystals and nets and the potentiation of skeletal muscle acto-SF1 ATPase activity at low ATP concentration. Likewise, earthworm tropomyosin inhibited skeletal muscle acto-HMM ATPase activity at normal ATP concentrations but to a much greater extent than skeletal muscle tropomyosin; this inhibition was removed by skeletal muscle troponin, in the presence of Ca2+. In a system containing earthworm myosin and skeletal muscle actin, earthworm tropomyosin had no detectable influence on the actin-activated ATPase activity. It is concluded that earthworm tropomyosin plays an active role in the actin-linked troponin-dependent regulatory system and has no measurable effect on the regulation via myosin.
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PMID:Properties of tropomyosin from the dual-regulated obliquely striated body wall muscle of the earthworm (Lumbricus terrestris L.). 621 Jul 9


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