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Biomedical subjects

M Rance

Publications and source records attributed to M Rance.

At least 19 recordsLinked to original sources
PubMed2005

Peak leg muscle power, peak VO2 and its correlates with physical activity in 57 to 70-year-old women.

The two aims of this study were first to measure short-term muscle power (STMP) by means of a cycling force-velocity test (cycling peak power: CPP) and a vertical jump test (jumping peak performance: JPP) and second, to examine the relationships between physical activity (PA) level, peak oxygen uptake (peak VO2) and STMP in healthy elderly women. Twenty-three independent community-dwelling elderly women (mean age: 64+/-4.4) performed on separate days, a peak oxygen uptake test on cycle ergometer, a cycling force-velocity test and a vertical jump test. A questionnaire (QUANTAP) was used to assess lifespan exercise habits. Four indices expressed in kJ day(-1) kg(-1) were calculated. Two indices represented average past PA level: 1/quantity of habitual physical activity (QHPA), 2/quantity of sports activities (QSA). Two indices represented the actual PA level: 3/actual quantity of habitual physical activity (AQHPA), 4/actual quantity of sports activities (AQSA). CPP (6.3+/-1.2 W kg(-1)) was closely correlated to JPP (14.8+/-3.4 cm) (r=0.80, P<0.001). AQHPA and AQSA were only positively associated with peak VO2 (ml min(-1) kg(-1)) (r=0.49; r=0.50, P<0.05, respectively). Past PA level was not related to fitness measurements. Results show that in this population: (1) jumping peak performance was closely related to CPP measured in the laboratory; (2) the cardio-respiratory fitness was related to the actual habitual physical activity level; (3) only age and anthropometric variables explained the actual performances in multivariate analysis.

Aged↗
PubMed2005

Alendronate-induced chemical laryngitis.

Alendronate is a bisphosphonate used in treating osteoporosis. Its recognized side-effects include oesophageal irritation and ulceration. The authors describe a case of laryngitis induced by transient contact of this medication with the laryngeal mucosa. Successful management of this case is also detailed.

Aged↗
PubMed2000

Regulatory domain conformational exchange and linker region flexibility in cardiac troponin C bound to cardiac troponin I.

Previously, we utilized (15)N transverse relaxation rates to demonstrate significant mobility in the linker region and conformational exchange in the regulatory domain of Ca(2+)-saturated cardiac troponin C bound to the isolated N-domain of cardiac troponin I (Gaponenko, V., Abusamhadneh, E., Abbott, M. B., Finley, N., Gasmi-Seabrook, G., Solaro, R.J., Rance, M., and Rosevear, P.R. (1999) J. Biol. Chem. 274, 16681-16684). Here we show a large decrease in cardiac troponin C linker flexibility, corresponding to residues 85-93, when bound to intact cardiac troponin I. The addition of 2 m urea to the intact cardiac troponin I-troponin C complex significantly increased linker flexibility. Conformational changes in the regulatory domain of cardiac troponin C were monitored in complexes with troponin I-(1-211), troponin I-(33-211), troponin I-(1-80) and bisphosphorylated troponin I-(1-80). The cardiac specific N terminus, residues 1-32, and the C-domain, residues 81-211, of troponin I are both capable of inducing conformational changes in the troponin C regulatory domain. Phosphorylation of the cardiac specific N terminus reversed its effects on the regulatory domain. These studies provide the first evidence that the cardiac specific N terminus can modulate the function of troponin C by altering the conformational equilibrium of the regulatory domain.

Animals↗
PubMed2000

Two-dimensional relayed-rotating-frame overhauser spectroscopy (1)H NMR experiments for the selective identification of 1,2-glycosidic linkages in polysaccharides.

This communication describes the use of two-dimensional relayed (TOCSY)-ROESY experiments for the rapid and selective identification of alpha/beta1,2-glycosidic linkages in polysaccharides. The method assists in the identification of cross-peaks in crowded regions of ROESY spectra by moving them to less congested areas. In addition, the appearance of the spectra provides information relating the location of the glycosidic linkage within the sequence of the glycan under study. Selection of solely the 1,2- linkages is achieved by appropriately tuning the duration of the TOCSY mixing period. The method is demonstrated both theoretically and experimentally for a variety of test case polysaccharides.

Carbohydrate Sequence↗
PubMed2000

Characterization of the binding interface between the E-domain of Staphylococcal protein A and an antibody Fv-fragment.

Staphylococcal protein A (SpA) is a cell-surface component of Staphylococcus aureus. In addition to the well-characterized interaction between SpA and the Fc-region of human IgG, an alternative binding interaction between SpA and the Fab-region of immunoglobulin domains encoded by the V(H)3 gene family has been described. To characterize structurally the interface formed by SpA repeats and type-3 V(H)-domains, we have studied the 32-kDa complex formed between an E-domain mutant (EZ4) and the Fv-fragment of the humanized anti-HER2 antibody (Hu4D5-8) using heteronuclear NMR spectroscopy. Protocols were established for efficient incorporation of (15)N, (13)C, and (2)H into EZ4 and the V(H)- and V(L)-domains of the Fv, allowing backbone resonances to be assigned sequentially for EZ4 and the V(H)-domain in both free and complexed states. Broadening of certain V(H)-resonances in the free and bound Fv-fragment suggests microsecond to millisecond time-scale motion in CDR3. Residues experiencing significant chemical shift changes of backbone (1)H(N), (15)N, and (13)CO resonances upon complex formation delineate contiguous surfaces on EZ4 and the V(H)-domain that define the binding interfaces of the two proteins. The interaction surfaces identified by chemical shift mapping are comprised of predominantly hydrophilic residues. This is in contrast to the SpA-Fc interface which is predominantly hydrophobic in nature. Further analysis of the surface properties suggests a probable binding orientation for SpA- and V(H)3-domains.

Amino Acid Sequence↗
PubMed2000

Site-site communication in the EF-hand Ca2+-binding protein calbindin D9k.

The cooperative binding of Ca2+ ions is an essential functional property of the EF-hand family of Ca2+-binding proteins. To understand how these proteins function, it is essential to characterize intermediate binding states in addition to the apo- and holo-proteins. The three-dimensional solution structure and fast time scale internal motional dynamics of the backbone have been determined for the half-saturated state of the N56A mutant of calbindin D9k with Ca2+ bound only in the N-terminal site. The extent of conformational reorganization and a loss of flexibility in the C-terminal EF-hand upon binding of an ion in the N-terminal EF-hand provide clear evidence of the importance of site-site interactions in this family of proteins, and demonstrates the strength of long-range effects in the cooperative EF-hand Ca2+-binding domain.

Allosteric Site↗
PubMed1999

NMR analysis of cardiac troponin C-troponin I complexes: effects of phosphorylation.

Phosphorylation of the cardiac specific amino-terminus of troponin I has been demonstrated to reduce the Ca2+ affinity of the cardiac troponin C regulatory site. Recombinant N-terminal cardiac troponin I proteins, cardiac troponin I(33-80), cardiac troponin I(1-80), cardiac troponin I(1-80)DD and cardiac troponin I(1-80)pp, phosphorylated by protein kinase A, were used to form stable binary complexes with recombinant cardiac troponin C. Cardiac troponin I(1-80)DD, having phosphorylated Ser residues mutated to Asp, provided a stable mimetic of the phosphorylated state. In all complexes, the N-terminal domain of cardiac troponin I primarily makes contact with the C-terminal domain of cardiac troponin C. The nonphosphorylated cardiac specific amino-terminus, cardiac troponin I(1-80), was found to make additional interactions with the N-terminal domain of cardiac troponin C.

Amino Acid Sequence↗
PubMed1999

Effects of troponin I phosphorylation on conformational exchange in the regulatory domain of cardiac troponin C.

Conformational exchange has been demonstrated within the regulatory domain of calcium-saturated cardiac troponin C when bound to the NH2-terminal domain of cardiac troponin I-(1-80), and cardiac troponin I-(1-80)DD, having serine residues 23 and 24 mutated to aspartate to mimic the phosphorylated form of the protein. Binding of cardiac troponin I-(1-80) decreases conformational exchange for residues 29, 32, and 34. Comparison of average transverse cross correlation rates show that both the NH2- and COOH-terminal domains of cardiac troponin C tumble with similar correlation times when bound to cardiac troponin I-(1-80). In contrast, the NH2- and COOH-terminal domains in free cardiac troponin C and cardiac troponin C bound cardiac troponin I-(1-80)DD tumble independently. These results suggest that the nonphosphorylated cardiac specific NH2 terminus of cardiac troponin I interacts with the NH2-terminal domain of cardiac troponin C.

Aspartic Acid↗
PubMed1999

Sensitivity improvement of transverse relaxation-optimized spectroscopy.

Procedures are described for significantly improving the sensitivity of the recently proposed TROSY (transverse relaxation-optimized spectroscopy) experiment (K. Pervushin et al., 1997, Proc. Natl. Acad. Sci. USA 94, 12366-12371). The TROSY experiment takes advantage of destructive interference between dipolar and chemical shift anisotropy relaxation mechanisms to achieve substantial reductions in resonance linewidths in heteronuclear correlation spectra; the effect is significant particularly for studies of large molecular weight systems at very high static magnetic field strengths. A (square root 2) improvement in the sensitivity of the TROSY experiment is achieved by implementation of the PEP (preservation of equivalent pathways) scheme (J. Cavanagh and M. Rance, 1990, J. Magn. Reson. 88, 72-85). An additional significant improvement in sensitivity for 15N-labeled samples in H2O solution is realized through a simple modification of the 1H-15N TROSY pulse sequence to return the water magnetization to its equilibrium position (+z axis) at the beginning of the acquisition period. Relaxation-induced imbalance between the coherence transfer pathways utilized in the TROSY refocusing period is shown theoretically and experimentally to give rise to additional unanticipated signals in TROSY spectra.

Calbindins↗
PubMed1999

Selective injection of magnetization by slow chemical exchange in NMR.

In a system in slow dynamic equilibrium two NMR methods are shown to be suitable for injecting magnetization from one resonance to another by means of slow chemical exchange. The combined outputs of the methods may be employed to measure the value of the off-rate constant kappa(off) in the complex. The methods are implemented experimentally using the complex of molecules composed of the enzyme Escherichia coli dihydrofolate reductase (DHFR) and the ligand folate. In an equilibrium solution with DHFR, folate is known to undergo chemical exchange between a free state and a bound state. The modified synchronous nutation method is applied to a spin of the folate molecule in the free and bound states; magnetization transfer occurs between the two sites due to the underlying exchange process. As a preliminary step for the application of the synchronous nutation method, a new one-dimensional 1H NMR technique is proposed which facilitates the assignment of the resonance of a spin in the bound state, provided the resonance of its exchange partner in the free state is known. This experiment is also used to obtain quantitative estimates of the transverse relaxation rate constant of the bound resonance. The numerical procedure necessary to analyze the experimental results of the synchronous nutation experiment is presented.

Escherichia coli↗
PubMed1999

Transverse-relaxation-optimized (TROSY) gradient-enhanced triple-resonance NMR spectroscopy.

Two modifications to sensitivity-enhanced gradient-selected TROSY-based triple-resonance NMR experiments are proposed that reduce the overall duration of the pulse sequences and minimize radiation damping effects on water-flipback solvent suppression. The modifications are illustrated for the HNCO-TROSY experiment, but are applicable to all triple-resonance experiments that detect proton magnetization after a reverse polarization transfer step from a (15)N spin. The methods are applied to yeast triosephosphate isomerase, a symmetric dimer with 248 amino acid residues per monomer.

Magnetic Resonance Spectroscopy↗
PubMed1999

A TROSY CPMG sequence for characterizing chemical exchange in large proteins.

A new NMR spin relaxation experiment is described for measuring chemical exchange time constants from approximately 0.5 ms to 5 ms in 15N-labeled macromolecules. The pulse sequence is based on the Carr-Purcell-Meiboom-Gill technique [Carr and Purcell (1954) Phys. Rev., 94, 630-638; Meiboom and Gill (1958) Rev. Sci. Instrum., 29, 688-691; Loria et al. (1999) J. Am. Chem. Soc., 121, 2331-2332], but implements TROSY selection [Pervushin et al. (1997) Proc. Natl. Acad. Sci. USA, 94, 12366-12371] to permit measurement of exchange linebroadening contributions to the narrower component of the 1H-15N scalar-coupled doublet. This modification extends the size limitation imposed on relaxation measurements due to the fast decay of transverse magnetization in larger macromolecules. The new TROSY-CPMG experiment is demonstrated on a [U-98% 15N] labeled sample of basic pancreatic trypsin inhibitor and a [U-83% 2H, U-98% 15N] labeled sample of triosephosphate isomerase, a 54 kDa homodimeric protein.

Binding Sites↗
PubMed1998

Adiabatic mixing in the liquid state.

Adiabatic spin inversion has been used in the liquid state very efficiently for decoupling purposes. Here we show that it can also be adapted for spin mixing experiments, such as the TOCSY and clean TOCSY experiment, and is superior to previously employed mixing sequences. The main advantage of adiabatic mixing sequences over the conventional mixing schemes used in liquid state experiments is an extremely low sensitivity to RF field inhomogeneity and miscalibration of the B1 field strength. The method is evaluated experimentally by comparing results obtained with different mixing schemes in the basic 2D TOCSY experiment. In addition to higher reliability, adiabatic mixing provides a sensitivity improvement of ca. 20% as compared to conventional mixing schemes. This is explained by higher signal losses due to RF inhomogeneity in the experiments employing traditional mixing schemes. More significant sensitivity improvements can be expected in situations where RF homogeneity is traditionally poor, for example, in large volume probes and magnetic resonance imaging experiments.

Algorithms↗
PubMed1997

High-resolution NMR structure and backbone dynamics of the Bacillus subtilis response regulator, Spo0F: implications for phosphorylation and molecular recognition.

NMR has been employed for structural and dynamic studies of the bacterial response regulator, Spo0F. This 124-residue protein is an essential component of the sporulation phosphorelay signal transduction pathway in Bacillus subtilis. Three-dimensional 1H, 15N, and 13C experiments have been used to obtain full side chain assignments and the 1511 distance, 121 dihedral angle, and 80 hydrogen bonding restraints required for generating a family of structures (14 restraints per residue). The structures give a well-defined (alpha/beta)5 fold for residues 4-120 with average rms deviations of 0.59 A for backbone heavy atoms and 1.02 A for all heavy atoms. Analyses of backbone 15N relaxation measurements demonstrate relative rigidity in most regions of regular secondary structure with a generalized order parameter (S2) of 0.9 +/- 0.05 and a rotational correlation time (taum) of 7.0 +/- 0.5 ns. Loop regions near the site of phosphorylation have higher than average rms deviation values and T1/T2 ratios suggesting significant internal motion or chemical exchange at these sites. Additionally, multiple conformers are observed for the beta4-alpha4 loop and beta-strand 5 region. These conformers may be related to structural changes associated with phosphorylation and also indicative of the propensity this recognition surface has for differential protein interactions. Comparison of Spo0F structural features to those of other response regulators reveals subtle differences in the orientations of secondary structure in the putative recognition surfaces and the relative charge distribution of residues surrounding the site of phosphorylation. These may be important in providing specificity for protein-protein interactions and for determining the lifetimes of the phosphorylated state.

Bacillus subtilis↗
PubMed1997

Rotational diffusion anisotropy of proteins from simultaneous analysis of 15N and 13C alpha nuclear spin relaxation.

Current methods of determining the rotational diffusion tensors of proteins in solution by NMR spectroscopy exclusively utilize relaxation rate constants for backbone amide 15N spins. However, the distributions of orientations of N-H bond vectors are not isotropic in many proteins, and correlations between bond vector orientations reduce the accuracy and precision of rotational diffusion tensors extracted from 15N spin relaxation data. The inclusion of both 13C alpha and 15N spin relaxation rate constants increases the robustness of the diffusion tensor analysis because the orientations of the C alpha-H alpha bond vectors differ from the orientations of the N-H bond vectors. Theoretical and experimental results for calbindin D9k, granulocyte colony stimulating factor, and ubiquitin, three proteins with different distributions of N-H and C alpha-H alpha bond vectors, are used to illustrate the advantages of the simultaneous utilization of 13C alpha and 15N relaxation data.

Amino Acid Sequence↗