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)

A 26K fragment of troponin T, which was produced by endogenous proteases in rabbit skeletal muscle, was isolated by SE-Sephadex column chromatography. This fragment sensitized both superprecipitation and ATPase of actomyosin to calcium ions, to the same extent as troponin T. There was no difference in affinity for tropomyosin between this fragment and troponin T as examined by affinity chromatography. Amino acid analysis showed that this fragment consisted of residues Ala-46-Lys-259 of troponin T. The N-terminal 45 residues of troponin T, therefore, are not essential for the physiological action of troponin T. It was also observed that Ca2+-activated neutral protease digested troponin T into the 26K fragment in the native thin filament, while the protease digested troponin T in a different way in the reconstituted thin filament.
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PMID:A 26K fragment of troponin T from rabbit skeletal muscle. 674 10

Actin filaments, when partially decorated with troponin T-I complexes, can slide faster on myosin heads than those with no decoration. Purified troponin T-I complexes bind to actins, and inhibit the actin activated myosin adenosine 5'-triphosphatase activity completely when the molar ratio of troponin T-I complex to actin is increased to 1 to 1. Those actin filaments decorated with troponin T-I complexes up to 20 to 50% of their molar ratio exhibit enhancement of the velocity of sliding on myosins up to 20% compared to those without such decoration. As the molar ratio of decoration further increases, the sliding velocity decreases. These results are consistent with the observation that even if some of actin monomers do not participate in the ATPase activity directly, they can interact with myosin heads and take part in the sliding movement.
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PMID:Decorating actin filaments with troponin T-I complexes and acceleration of their sliding movement on myosin molecules. 764 91

The study of the functional effects of troponin isoform changes would be greatly aided by the development of a strategy permitting protein engineering and mutational analysis. To assess the role of troponin isoforms in regulating myofibrillar ATPase activity, we have expressed rat cardiac troponin I (cTnI) in E. coli and purified the protein to near homogeneity. We utilized the inducible expression vector pGEX-KG to create a glutathione-S-transferase fusion protein which can be cleaved with thrombin. Approximately 6 mg of cTnI can be purified from 1 l of culture. Ca2+Mg2+ ATPase activity was measured using the bacterially synthesized cTnI and the remaining components of the regulated actomyosin complex (troponin T, troponin C, tropomyosin, actin, and myosin) purified to homogeneity from mammalian hearts. In the presence of free Ca2+ ranging from 10(-2) to 10(-8) M, bacterially synthesized cTnI exhibits specific activity similar to that observed for control cTnI isolated from rat hearts. The bacterially synthesized protein is capable of stoichiometric phosphorylation and demonstrates appropriately regulated specific activity. These results establish the feasibility of using bacterial expression to study functional consequences of changes in expression of troponin isoforms.
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PMID:Expression of regulated cardiac troponin I in Escherichia coli. 773 Oct 51

Troponin in bullfrog skeletal myofibrils was substituted by troponin T and troponin T1 from rabbit skeletal muscle. The inhibitory effect of troponin I was examined on the ATPase activity of these troponin-substituted myofibrillar preparations. The ATPase of myofibrils substituted by troponin T was more inhibited by troponin I than the ATPase of myofibrils substituted by troponin T1.
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PMID:The inhibitory effect of troponin I on the ATPase activity of myofibrils treated with troponin T and troponin T1. 775 Aug 94

The M(r) 52,000 subunit of Akazara scallop striated muscle troponin, which was tentatively identified as troponin I, was cleaved into two major fragments with CNBr: C-terminal 17 kDa fragment (CN17K) and N-terminal 35 kDa fragment (CN35K) [J. Biochem. 108, 519-521 (1990)]. CN17K inhibits rabbit reconstituted actomyosin Mg-ATPase activity, weakly in the absence of troponin T but strongly in its presence, together with Akazara tropomyosin. CN35K, however, hardly shows such inhibition. Thus, the amino acid sequence of the CN17K was determined by the Edman method. CN17K comprises 135 amino acid residues and its calculated molecular mass is 15,732 Da. A computer search of the SWISS-PROT data base revealed the TnIs of crayfish tail muscle, rabbit skeletal muscle, and bovine cardiac muscle to be homologous proteins with total sequence homologies of 39, 30, and 30%, respectively, to CN17K. Significantly high homology was observed among these TnIs in the regions around residues 75-95, 99-114, and 135-151 of the rabbit TnI. From these facts, we conclude that the 52K subunit is a TnI.
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PMID:Amino acid sequence of C-terminal 17 kDa CNBr-fragment of Akazara scallop troponin-I. 777 83

The myocardium is a highly adaptive tissue, as evidenced by phenotypic alterations throughout development and under conditions of altered hemodynamic load. With pressure overload, the myocardium displays adult-to-fetal transitions in expression of contractile and non-contractile proteins. Most intriguing is the fact that many of these transitions are also observed in the senescent heart. The purpose of this work was to establish if the thin filament regulatory proteins, troponin I and troponin T, exhibit reexpression of early developmental isoforms, suggestive of coordinate reprogramming of contractile protein isoform expression. As a functional index of reexpression of the early isoform of troponin I, slow skeletal troponin I, myofibrils were isolated from 12 and 24-month-old Fischer 344 rat ventricles and assayed for myofibrillar ATPase activity at pH 7.0 and 6.5. Both preparations displayed rightward shifts in Ca-ATPase relationships with no differences between groups. SDS-PAGE and Western blot analysis showed that whereas myosin heavy chain expression underwent a transition to predominance of the early development isoform, beta-myosin heavy chain, there was no reexpression of the fetal isoforms of either troponin I or troponin T in the rat heart at 24 months of age. Northern blot analysis using cDNA probes specific for cardiac or slow skeletal troponin I also confirmed the lack of slow skeletal reexpression in the 24-month ventricle. These results are significant in that they demonstrate a lack of coordinate expression of contractile protein isoforms under myocardial adaptation to the aging process.
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PMID:Discoordinate regulation of contractile protein gene expression in the senescent rat myocardium. 807 7

The effect of partial removal of troponin I and C on the profiles of Ca(2+)-sensitive ATPase activity in rabbit skeletal myofibrils was investigated by replacing the troponin C.I.T-complex in the myofibrils with exogenously added troponin T under the same conditions as those reported previously [Shiraishi et al. (1992) J. Biochem. 111, 61-65]. During the course of the troponin T treatment, the level of the ATP hydrolysis at low Ca2+ concentrations was elevated and the pCa for half maximum activation increased, while the cooperativity decreased. These changes in the parameters of the ATPase were correlated with the extent of the troponin I removal from myofibrils.
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PMID:The effect of partial removal of troponin I and C on the Ca(2+)-sensitive ATPase activity of rabbit skeletal myofibrils. 818 28

Three subunits of rabbit skeletal muscle troponin were expressed in and purified from Escherichia coli. The procedures were optimized, and the reconstituted troponin complex is highly homogeneous, stable, and obtainable in large quantities, allowing us to conduct crystallization studies of the troponin complex. The three subunits expressed and purified are beta-TnT(N'-208), TnI(C64A, C133S), and the wild type TnC. beta-TnT(N'-208) is a 25 kDa fragment of beta-troponin T, which consists of 208 amino acids and lacks 58 residues in the N-terminal variable region. TnI(C64A, C133S) is a mutant troponin I, in which Cys-64 and Cys-133 are replaced by Ala and Ser, respectively. Each subunit was separately expressed in E. coli, purified by column chromatography including HPLC, and reassembled to form troponin complex. The reconstituted troponin complex was not distinguishable from authentic troponin prepared from rabbit skeletal muscle; the acto-S1 ATPase rate, as well as the superprecipitation, was calcium-sensitive. Small flat crystals up to 0.2 mm long have been reproducibly obtained in preliminary crystallization trials.
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PMID:Reconstitution of rabbit skeletal muscle troponin from the recombinant subunits all expressed in and purified from E. coli. 828 38

The Ca(2+)-regulatory proteins, i. e., troponin C, I and T, were prepared from puffer skeletal muscle. The SDS-gel electrophoretic study indicated that the molecular weights of troponin C, I and T were 18, 20 and 30k daltons, respectively. The Ca(2+)-sensitivity of the ATPase activity of the puffer myofibrils was abolished by the removal of troponin C by the CDTA-treatment and then restored by the reconstitution with puffer skeletal troponin C. The treatment with excess troponin T of either puffer or rabbit skeletal muscle at slightly acidic condition resulted in the removal of troponins C and I from the puffer myofibrils. The reconstituted puffer troponin C-I-T gave the Ca(2+)-sensitivity to the ATPase activity of puffer desensitized myofibrils in the presence tropomyosin.
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PMID:Preparation of three troponin components from puffer skeletal muscle. 835 80

Familial hypertrophic cardiomyopathy (HCM) can be caused by dominant missense mutations in cardiac troponin T (TnT), alpha-tropomyosin, C-protein, or cardiac myosin heavy chain genes. The myosin mutations are known to impair function, but any functional consequences of the TnT mutations are unknown. This report describes the in vitro function of troponin containing an IIe91Asn mutation in rat cardiac TnT, corresponding to the HCM-causing Ile79Asn mutation in man. Mutant and wild-type TnT cDNAs were expressed in bacteria and the proteins purified and reconstituted with the other troponin subunits, the mutation had no effect on troponin's affinity for tropomyosin, troponin-induced binding of tropomyosin to actin, cooperative binding of myosin subfragment 1 to the thin filament, CA(2+)-sensitive regulation of thin filament-myosin subfragment 1 ATPase activity, or the CA2+ concentration dependence of this regulation. However, the mutation resulted in 50% faster thin filament movement over a surface coated with heavy meromyosin in in vitro motility assays. The increased sliding speed suggests an unexpected role for the amino terminal region of TnT in which this mutation occurs. The relationship between this faster motility and altered cardiac contraction in patients with HCM is discussed.
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PMID:Altered cardiac troponin T in vitro function in the presence of a mutation implicated in familial hypertrophic cardiomyopathy. 867 96


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