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

The extrusion kinetics of two cruciforms derived from unrelated DNA sequences differ markedly. Kinetic barriers exist for both reactions, necessitating elevated temperatures before extrusion proceeds at measureable speeds, but the dependence upon temperature and ionic strength is quite different for the two sequences. One, the ColE1 inverted repeat, exhibits a remarkably great temperature dependence of reaction rate and is suppressed by moderate amounts of NaCl or MgCl2. In contrast, the other, a synthetic inverted repeat present in pIRbke8, shows more modest temperature dependence and has a requirement for the presence of salt, with optimal concentrations being 50 mM NaCl or 100 microM MgCl2. Under optimal conditions, cruciform extrusion rates are fast (t1/2 less than 60m) at 37 degrees C for both sequences at native superhelix densities. In 50 mM NaCl the pIRbke8 inverted repeat is characterised by an Arrhenius activation energy of 42.4 +/- 3.2 kcal mole -1. The differences in kinetic properties between the two sequences indicate that DNA base sequence is itself an important factor in determining cruciform kinetics, and possibly even in the selection of the mechanistic pathway.
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PMID:The kinetic properties of cruciform extrusion are determined by DNA base-sequence. 400 Sep 40

The thermodynamic parameters characterizing the interaction between rabbit fast skeletal muscle troponin and tropomyosin have been determined at 25 degrees C for three solution conditions: buffer containing (A) 1 mM CaCl2, simulating a "turned-on" state, (B) 3 mM MgCl2, simulating a "turned-off" state, and (C) 2 mM ethylenediaminetetraacetic acid, a reference state. The enthalpies were measured in two buffers with different heats of ionization to allow correction for dissociation or uptake of protons. The enthalpies corrected for proton effects are -22.1, -25.4, and -23.5 kcal/mol, respectively, in buffers A, B, and C. The interaction between troponin and tropomyosin in the presence of calcium is accompanied by release of 0.9 mol of proton per mole of complex. Proton effects in the presence of magnesium and in the absence of divalent metal ions were too small to quantitate. The association constants were measured by using tropomyosin labeled with the extrinsic fluorescent probe dansylaziridine, and binding was detected by enhancement of the probe fluorescence. The magnitudes of the association constants for unlabeled troponin are 7.5 X 10(5), 4.2 X 10(5), and 9.5 X 10(5) M-1, respectively, for the three solution conditions corresponding to unitary free energies of -10.4, -10.1, and -10.6 kcal/mol. The unitary entropies for the interaction are -39, -51, and -43 cal/(deg X mol), respectively, for the three solution conditions. Under these conditions, the troponin-tropomyosin interaction is enthalpy driven, and a large unfavorable entropy must be overcome in the formation of the complex.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Interaction of troponin and tropomyosin: spectroscopic and calorimetric studies. 407 90

Incubation of inorganic pyrophosphate from baker's yeast with phosphate and MgCl2 in the presence of fluoride results in a gradual inactivation of the enzyme concomitant with incorporation of PP1 (about 2 moles per mole) into the protein. The rate constant for this process shows an increase with a rise in concentrations of the three reagents, the maximal value of inactivation being 0.11 min-1. The bound PP1 is not separated by gel-filtration. The rate of spontaneous degradation of the enzyme-pyrophosphate complex and the nature of EDTA and Mg2+ effects are similar to those for the analogous compound obtained by inhibition of PP1 hydrolysis by fluoride. The data obtained suggest that during PP1 synthesis and hydrolysis by pyrophosphatase fluoride stabilizes the same intermediate of the enzyme with pyrophosphate.
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PMID:[Stable compound of inorganic pyrophosphatase with pyrophosphate obtained by a fluoride-mediated reaction with phosphate]. 611 33

Two different types of essential carboxyl groups were detected in the extrinsic component of the proton ATPase of Rhodospirillum rubrum. Chemical modification of R. rubrum chromatophores or its solubilized ATPase by Woodward's reagent K resulted in inactivation of photophosphorylating and ATPase activities. The apparent order of reaction was nearly 1 with respect to reagent concentration and similar K1 were obtained for the soluble and membrane-bound ATPases suggesting that inactivation was associated with modification of one essential carboxyl group located in the soluble component of the proton ATPase. Inactivation was prevented by adenine nucleotides but not by divalent cations. Dicyclohexylcarbodiimide completely inhibited the solubilized ATPase with a K1 of 5.2 mM and a K2 of 0.81 min-1. Mg2+ afforded nearly complete protection with a Kd of 2.8 mM. Two moles of [14C]dicyclohexylcarbodiimide were incorporated per mole of enzyme for complete inactivation but in the presence of 30 mM MgCl2 only one mole was incorporated and there was no inhibition. The labeling was recovered mostly from the beta subunit. The incorporation of the labeled reagent into the ATPase was not prevented by previous modification with Woodward's reagent K. It is concluded that both reagents modified two different essential carboxyl groups in the soluble ATPase from R. rubrum.
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PMID:Two types of essential carboxyl groups in Rhodospirillum rubrum proton ATPase. 630 54

Glutamyl-tRNA synthetase has been isolated from an extreme thermophile, Thermus thermophilus HB8. The enzyme has been purified to homogeneity by successive chromatography on columns of DEAE-cellulose, DEAE-Sephacel, phosphocellulose and hydroxyapatite. 11.7 mg of purified enzyme has been obtained from 2 kg of T. thermophilus cells, with a purification factor of 600 with an 11% yield. From gel permeation chromatography and sodium dodecyl sulfate polyacrylamide gel electrophoresis, the enzyme is found to be a monomer protein with a molecular weight of 50,000. The optimum temperature for the aminoacylation of T. thermophilus tRNAGlu is 65 degrees C, and the optimum pH range is 8.0-9.0, in the presence of 5 mM Mg2+. The Km values for ATP, L-glutamate, and T. thermophilus tRNAGlu are 230 microM, 70 microM, and 0.65 microM, respectively, in the presence of 50 mM KCl and 10 mM MgCl2 at pH 8.0 at 65 degrees C. Escherichia coli tRNA2Glu is also a good substrate with a Km value of 0.60 microM at 65 degrees C. The mole fractions of Arg and Leu residues are higher and that of Asx residues is lower than those of E. coli glutamyl-tRNA synthetase. Glutamyl-tRNA synthetase from T. thermophilus is remarkably thermostable; even after incubation for 9 h at 65 degrees C, 70% of the enzyme activity is retained in the absence of any protecting factors. Such an extremely thermostable enzyme with a low molecular weight will be useful for detailed physiochemical analyses on the molecular mechanism of strict recognition by aminoacyl-tRNA synthetases.
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PMID:Purification and characterization of glutamyl-tRNA synthetase from an extreme thermophile, Thermus thermophilus HB8. 652 23

The experimental conditions for release of the regulatory light chain (RLC) of scallop myosin at 30 degrees C were studied. Substantially all RLC was released from myosin by incubation for 5 min in medium containing buffer and KCl. This release of RLC was inhibited strongly by Ca2+, while the effect of Mg2+ was about 10,000 times weaker than that of Ca2+. Even in the absence of Ca2+, MgATP and MgADP inhibited the release of RLC, while the protective effect of AMPPNP was negligible. Other Mg nucleotides also showed some protective effect, though appreciably less than MgATP. The incubation of scallop myosin with abalone regulatory light chain (LC2) at 30 degrees C for 5 min produced a hybrid myosin. In the presence of 5 mM MgCl2, 1 of the 2 mol of RLC per mol of scallop myosin was exchanged with 1 mol of LC2. In the presence of Ca2+ or MgATP, myosin bound 1 extra mole of LC2 besides the 2 mol each of SH-LC and RLC.
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PMID:Inhibitory effect of MgATP on the release of regulatory light chain from scallop myosin and light chain composition of scallop myosin hybridized with abalone light chain 2 at 30 degrees C. 660 40

The structure of the major protein constituent of photosynthetic membranes in higher plants, the chlorophyll a/b-light harvesting complex (LHC), was studied by x-ray diffraction and electron microscopy. The LHC was purified from Triton X-100 solubilized thylakoid membranes of the pea, and contained 6 mol of chlorophylls a and b per mole of a polypeptide of 27,000 molecular weight. X-ray diffraction showed that in the presence of 10 mM MgCl2, purified LHC forms planar aggregates that stack with a period of 51 A. Within each layer, LHC molecules pack with a center-to-center distance of 85 A but without long-range order. However, when LHC is incorporated into single-walled vesicles of plant lecithin, the addition of NaCl above 10 mM, or MgCl2 above 2 mM, led to the formation of plaques of hexagonal lattices, with a lattice constant of 125 A. The large domain size and high degree of order in the plane of the membrane are evident from the sharp lattice lines observed to 7 A resolution on the equator of the x-ray pattern. Freeze-fracture electron micrographs demonstrated an aligned stacking of the lattices in adjacent membranes, resulting in crystallinity in the third dimension over short distances. Micrographs of negatively stained membranes revealed a hexagonal lattice of the same lattice constant, formed by surface-exposed parts of the LHC molecules which are probably responsible for the ordered stacking of lattices. In both the LHC aggregates and in the reconstituted membrane lattices the diffracted x-ray intensities at 10-A spacing on the equator indicate that the LHC molecule contains paralled alpha-helices or beta-sheets that are oriented perpendicular to the planar arrays.
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PMID:Formation of crystalline arrays of chlorophyll a/b - light-harvesting protein by membrane reconstitution. 703 99

Using stepwise protein fractionation by (NH4)2SO4 and ion-exchange chromatography on CM-cellulose, DEAE-cellulose and SP-Sephadex, two isoforms of 6-phosphogluconate dehydrogenase, A and B, from rat liver were obtained. The method developed allows to achieve complete separation of these forms and to obtain preparative amounts of the protein with specific activities of 5.7 and 10.7, respectively. The native enzyme forms A and B have molecular weights of 107000 and are represented by dimers composed of subunits with identical molecular weights equal to 54000. Both isoforms have a pH optimum at 8.5 and reveal different sensitivity to MgCl2 and MnCl2. The tetrahedron-shaped ions (phosphate, molibdate, arsenate) inhibit the both molecular forms of the enzyme, The Arrhenius plots for the reaction rate are uninterrupted lines within the temperature range of 21-44 degrees; the values of activation energy and the temperature coefficient for isoforms A and B are 12750 and 13500 cal/mole and 2.05 and 2.15, respectively.
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PMID:[Purification and some properties of molecular forms of 6-phosphogluconate dehydrogenase from rat liver]. 715 26

Soluble complexes of poly (U) and adenylic nucleotides in NaCl solutions were studied by scanning microcalorimetry. The melting enthalpies, delta Hm, of poly (U) complexes with adenosine, 2',3' -cAMP, 2'(3')-AMP, 5-AMP, ADP, ATP in 1 M NaCl are 50.5; 45.0; 42.9; 28.6; 26.1 and 25.6 kJ/mole triplets, respectively. Delta Hm is independent of the complex melting temperature, Tm. The calorimetric enthalpies are considerably lower than the apparent delta Hv.H. obtained from Tm dependence on free monomer concentration. The enthalpy of complex formation in 1 M NaCl depends neither ob the number nor on the degree of ionization of the phosphate groups but is essentially determined by their 5' - or 2'(3')-position. In contrast to 2'(3')- AMP. 2 poly (U), delta Hm of 5'AMP. 2 poly (U) increases considerably at lowering Na+ concentration. The enthalpy of poly (U) double helix melting in 1 M NaCl is 8.8 kJ/mole pairs which is 2.5 times lower than that in MgCl2 solutions.
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PMID:Calorimetric study of the complexes between polyuridylic acid and adenylic nucleotides. 730 78

Near-UV difference spectral analysis of the triplex formed from d(C-T)6 and d(A-G)6.d(C-T)6 in neutral and acidic solution shows that the third strand dC residues are protonated at pH 7.0, far above their intrinsic pKa. Additional support for ion-dipole interactions between the third strand dC residues and the G.C target base pairs comes from reduced positive dependence of triplet stability on ionic strength below 0.9 M Na+, inverse dependence above 0.9 M Na+ and strong positive dependence on hydrogen ion concentration. Molecular modeling (AMBER) of C:G.C and C+:G.C base triplets with the third strand base bound in the Hoogsteen geometry shows that only the C+:G.C triplet is energetically feasible. van't Hoff analysis of the melting of the triplex and target duplex shows that between pH 5.0 and 8.5 in 0.15 M NaCl/0.005 M MgCl2 the enthalpy of melting (delta H degree obs) varies from 5.7 to 6.6 kcal.mol-1 for the duplex in a duplex mixture and from 7.3 to 9.7 kcal.mol-1 for third strand dissociation in the triplex mixture. We have extended the condensation-screening theory of Manning to pH-dependent third strand binding. In this development we explicitly include the H+ contribution to the electrostatic free energy and obtain [formula: see text]. The number of protons released in the dissociation of the third strand from the target duplex at pH 7.0, delta n2, is thereby calculated to be 5.5, in good agreement with approximately six third strand dc residues per mole of triplex. This work shows that when third strand binding requires protonated residues that would otherwise be neutral, triplex formation and dissociation are mediated by proton uptake and release, i.e., a proton switch. As a by-product of this study, we have found that at low pH the Watson-Crick duplex d(A-G)6.d(C-T)6 undergoes a transition to a parallel Hoogsteen duplex d(A-G)6.d(C(+)-T)6.
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PMID:UV spectroscopic identification and thermodynamic analysis of protonated third strand deoxycytidine residues at neutrality in the triplex d(C(+)-T)6:[d(A-G)6.d(C-T)6]; evidence for a proton switch. 765 30


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