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Query: UMLS:C0027960 (mole)
 21,279 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Recent advances in the studies of the aggregation of G-actin monomers, containing one molecule of ATP, to long filaments of F-actin, with a concomitant hydrolysis of the nucleotide to ADP, are reviewed. With the aid of omega-ATP, the association and dissociation rate constant of the nucleotide could be determined. The binding of the nucleotide is enhanced by the binding of one Ca++ ion, probably at a different site. The delta G value of the Mg++ or Ca++ induced polymerization has been determined to --39 to--59 kJ/mole, the critical protein concentration for the ATP-G-actin to ADP-F-actin conversion is very strongly influenced by the concentration of bivalent cations. The rate constants of the protein monomers, and the rate and equilibrium constants for the propagation step show the process to be extremely cooperative. Actin shows the interesting phenomenon of translocational head-to-tail polymerization, which may be regulated by ATP. The contact sites between the monomers in F-actin have been labeled by chemical modification. Two tryosine residues, 53 and 69, are probably close to one of the two sites. The ATP binding sites has been labeled by an ATP analog, and there is evidence that it is close to the contact site.
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PMID:The polymerization reaction of muscle actin. 34 Sep 37

The binding of adenosine diphosphate-ribosylated elongation factor 2 (ADPRib-EF-2) to ribosomes was inhibited both in the presence and absence of GTP in proportion to the amounts of unmodified EF-2 added. Concomitant with this inhibition, an increase in the activity of ribosome-bound EF-2 in polyphenylalanine synthesis was observed. On the other hand, the addition of ADPRib-EF-2 reduced the rate of poly(Phe) synthesis observed in the presence of a saturating amount of EF-2 and increased the amount of EF-2 required for the half-maximal rate of poly(Phe) synthesis. Phe-tRNA, nonenzymatically bound to the ribosome in the presence of poly(U), inhibited the subsequent binding of ADPHRib-EF-2. The same ribosomal population appeared to preferentially bind either aminoacyl-tRNA or ADPRib-EF-2. The Scatchard plot of the binding of ADPRib-EF-2 to the ribosome in the presence of GTP revealed the presence of two ribosomal binding sites (or ribosomal populations) with apparent different affinities for the modified factor (K371 degrees d,1 = 6.6 nM and K37 degrees d,2 = 126 nM). At saturating concentrations of ADPRib-EF-2, a maximum of about 1 molecule of the factor was bound per ribosome. The binding of ADPRib-EF-2 to the ribosome was stimulated by GTP. The binding of radioactive GTP to the ribosome was observed concomitantly with the binding of ADPRib-EF-2. One mole of GTP was bound per mole of ADPRib-EF-2. No significant difference could be found in the binding of GTP to ribosome required in the presence of either EF-2 or ADPRib-EF-2. The binding of ADPRib-EF-2 to the ribosome required the presence of Mg2+ and reached a maximum at 5 mM. The binding was greatest at K+ concentrations below 20 mM. ADPRib-EF-2 was bound primarily to the large ribosomal subunit. A slight, but reproducible binding to the 40 S subunit was also observed. The addition of 40 S to 60 S subunits stimulated the binding of ADPRib-EF-2. GTP displayed a stimulatory effect on the binding only in the presence of recombined subunits. Human ADPRib-EF-2 was bound to rat liver ribosomes as efficiently as to human tonsil ribosomes, while the binding to Escherichia coli ribosomes was insignificant.
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PMID:Interactions of adenosine diphosphate-ribosylated elongation factor 2 with ribosomes. 78 67

Binding of the nucleotides ATP and ADP by preparations of sarcoplasmic reticulum was investigated by the method of flow dialysis. For ATP, experimental data could not be analyzed directly in terms of binding since a significant though small amount of hydrolysis could be observed even in presence of EDTA. ADP binding could be analyzed and gave a dissociation constant of 10-20 muM at neutral pH, and a stoichiometry of 0.35 - 0.45 per mole ATPase. The possible significance of this stoichiometry is discussed. Similar experiments were performed after ethoxyformylation of sarcoplasmic reticulum which inhibits the Ca2+ dependnet ATPase activity. The results confirmed the inhibition of ATP hydrolysis and pointed to a considerably reduced affinity for nucleotides. The method based on the measurement of dialysis rates is convenient, and accurate enough to detect the effects of chemical modification on sarcoplasmic reticulum membranes.
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PMID:Binding of nucleotides ATP and ADP to sarcoplasmic reticulm: study by rate of dialysis. 95 55

Subfragment-1 prepared by chymotryptic digestion of myosin was applied to a column of Sepharose-adipic acid hydrazide-ATP in 1 mM EDTA, 10 mM Tris-HCL (PH 7.6), and 40 mM KCL. Ninety-nine per cent of subfragment-1 was adsorbed on the column in this medium. Fourty-three per cent of the applied protein was eluted with 6 mM ADP in the above buffer and then 52% was eluted with 1 mM EDTA, 10 mM Tris-HCL (pH 7.6), AND 0.7 M KCL. The former fraction contained g3 chain and the latter g1 chain. These fractions were apparently the same as the components, p2 and p1, respectively, isolated by ion-exchange chromatography using DEAE-cellulose (Yagi & Otani (1974) J. Biochem. 76, 365-373). No significant difference of ADP binding was found between the two fractions, both could bind about 0.5 mole per 10(5) g of protein. The preparation of the two subfragment-1 fractions is described.
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PMID:Separation of myosin subfragment-1 into fractions containing g1 chani and g2 chain by Sepharose-adipic acid hydrazide-ATP column chromatography. 95 34

The interaction of diethylpyrocarbonate (DEP) with the pyruvate dehydrogenase component (PDH) isolated from the pyruvate dehydrogenase complex (EC 1.2.4.1) results in a modification of 3-5 histidine residues per mole of enzyme, which simultaneously decreases the enzyme activity. After PDH inhibilion by DEP in the presence of dithiothreitol almost complete reactivation (94%) under the effect of neutral hydroxylamine is observed. In the absence of SH-groups protection incomplete reactivation by hydroxylamine (79%) is found. In the latter case titration with 5,5-dithio--bis-(2-nitrobenzoic acid) in 8 M urea showed that the DEP-modified protein contains less quantity of SH groups (by 4-8) as compared to the native enzyme. It is assumed that the DEP-modified SH-groups are not responsible for the enzyme activity. The differential spectrum of the modified and native PDH showed no changes within the range of 260-300 nm. TPP in combination with Mg2+ (10(-3) M) protectes PDH from being inactivated by DEP. TPP (10(-2) M) reactivates PDH by 70% after its complete inhibition by DEP. Similar protective action is manifested by ATP, ADP and inorganic pyrophosphate in the presence of Mg2+. A kinetic study showed a competitive type of PDH inhibition by DEP with respect to TPP. it is concluded that the histidine residues of PDH are involved in TPP binding.
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PMID:[Role of muscle pyruvate dehydrogenase histidine residues in thiamine pyrophosphate binding]. 98 22

The effect of Mg2+ on the binding of adenylates to isolated chloroplast coupling factor 1 (CF1) was studied using CD spectrometry and ultrafiltration. At adenylate concentrations smaller than 100 muM, one mole of CF1 binds three moles of ATP (or ADP) regardless of the presence of Mg2+. In the presence of Mg2+, the first two ATP's bind to CF1 independently with the same binding constant of 2.5 X 10(-1) muM-1, then the third ATP binds with a much higher affinity of 10 muM-1. In the absence of Mg2+, the first ATP binds to CF1 with a binding constant of 2.5 X 10(-1) muM-1 then the other two ATP's bind less easily with the same binding constant of 4.0 X 10(-2) muM-1. The binding mode of ADP to CF1 is quite similar to that of ATP. In the presence of Mg2+, the binding constants of the first two ADP's are both 7.6 X 10(-2) muM-1, that of the third ADP being 4.0 muM-1. In the absence of Mg2+, the binding constant of the first ADP is 7.6 X 10(-2) muM-1, the constants of the other two ADP's both being 4.0 X 10(-2) muM-1. AMP caused a negligible change in CD.
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PMID:Magnesium ion-induced changes in the binding mode of adenylates to chloroplast coupling factor 1. 100 84

We report measurements of the reactivity (degree of labeling, as mole of ligand per mole of protein, at constant exposure time) of the reactive thiol, "SH1", of a subfragment of myosin (S-1), and of Cys-10 of F-actin under various conditions, using N-iodo-[3H]acetyl-N-(1-sulfo-5-naphthyl)ethylenediamine, a fluorescent radioactive iodoacetamide analog. When either ADP or adenyloyl imidodiphosphate (simulating unhydrolyzed ATP) is bound to the enzymatic site of S-1, the reactivity of "SH1" is slightly enhanced, but when active ATPase is going on, reactivity is reduced by about a third, presumably due to the species, (S-1) ADP,Pi. The reactivity of Cys-10 alone is very low. When the complex, (S-1)-F-actin, is formed, the reactivity of SH1 is strongly decreased, and the reactivity of Cys-10 is strongly increased. The foregoing results explain our further observation (on glycerol-treated rabbit psoas fibers) that when fibers labeled in relaxation solution are compared with fibers labeled in rigor solution, myosin is more reactive and actin is less reactive, in the former case; alpha-actinin and C-protein are also less reactive in the former case.
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PMID:Reciprocal reactivities of specific thiols when actin binds to myosin. 106 Nov 33

The binding of ADP to heavy meromyosin has been studied by microcalorimetry. Minute amounts of myokinase interfere with binding measurements, but by selection of appropriate conditions, we can estimate that the value of the apparent deltaHbinding lies between - 1.0 and - 3.0 kcal per mole of ADP bound (0.3 M KC1, 2 mM MgC12, 20mM Tris, pH 8.00, 20 degrees C). Values of deltaHbinding reported to date are an order of magnitude larger, and we suggest that these values are artifactual results due to myokinase contamination.
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PMID:On the enthalpy of binding of ADP to heavy meromyosin. 110 6

Platelets from individuals with familial hypercholesterolemia show increased sensitivity to the aggregating atents, epinephrine and ADP. Since the mechanism of this abnormal sensitivity is unknown, we examined, in vitro, the influence of the plasma lipid environment on the function of platelets. The composition of plasma lipids was altered by the addition of sonicated cholesterol-dipalmitoyl lecithin liposomes which were "cholesterol normal" (cholesterol-phospholipid mole ratio [C/P] equals 1.0, "cholesterol rich" (C/P eauals 2.2), or "cholesterol poor" (C/P equals 0). Cholesterol-normal liposomes had no influence on platelet lipids or platelet function. In contrast, after incubation for 5 h at 37 degrees C with cholesterol-rich liposomes, normal platelets acquired 39.2% excess cholesterol with no change in phospholipids or protein. The percent increase in platelet membrane cholesterol was three-fold that of the granule fraction. The acquisition of cholesterol by platelets was associated with a 35-fold increase in sensitivity to epinephrine-induced aggregation (P less than 0.001) and 15-fold increase to ADP aggregation (P less than 0.001), as determined both by aggregometry and by [13C]serotonin release. Response to thrombin or collagen was unchanged. Platelets incubated with cholesterol-poor liposomes underwent a selective loss of 21.4% cholesterol and this was associated with an 18-fold reduction in their sensitivity to epinephrine. These studies demonstrate that the cholesterol content of platelets is dependent on the lipid composition of the milier. Cholesterol acquired by platelets may exert its effect on platelet function by a modification of the platelet membrane.
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PMID:Platelet hypersensitivity induced by cholesterol incorporation. 111 69

A study of the equilibrium binding of ADP, 1,N6-ethenoadenosine diphosphate, adenylyl imidodiphosphate, and 1,N6-ethenoadenylyl imidodiphosphate to solubilized spinach chloroplast coupling factor 1 (CF1) has been carried out. All four nucleotides were found to bind to two apparently identical "tight" sites, with characteristic dissociation contants generally less than 10 muM. The binding to these "tight" sites is similar in the presence of Mg2+ and Ca2+, is stronger in 0.1 M NaC1 than in 20 mM Tris-C1, and is only slightly altered by heat activation. The slow rate of association of ADP and 1,N6-ethenoadenosine diphosphate at these sites rules out the possibility that they are catalytic sites for ATPase activity on the solubilized enzyme. A third tight site for adenylyl imidodiphosphate was found on the heat-activated enzyme. The dissociation constant for this interaction (7.6 muM) is similar to the adenylyl imidodiphosphate competitive inhibition constant for ATPase activity (4 muM). ADP, which inhibits ATPase activity but is not a strong competitive inhibitor, binds only weakly at a third site (dissociation constant greater than 70 muM). One mole of 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole reacted per mole of CF1 prevents ADP and adenylyl imidodiphosphate binding at the "catalytic" site and abolishes the ATPase activity. A model is proposed in which the "tight" nucleotide binding sites act as allosteric conformational switches for the ATPase activity of solubilizedCF1.
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PMID:Characterization of nucleotide binding sites on chloroplast coupling factor 1. 114 87


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