A Proton Magnetic Resonance Study of the Association of Lysozyme with Monosaccharide Inhibitors

It has been shown that the acetamido methyl protons of N-acetyl-d-glucosamine undergo a chemical shift to higher fields in their proton magnetic resonance spectrum when the inhibitor is bound to lysozyme. The observed chemical shift in the presence of the enzyme is different for the agr- and ß-anomeric forms of 2-acetamido-2-deoxy-d-glucopyranose indicating either a difference in the affinity of the anomeric forms for lysozyme or different magnetic environments for the methyl protons in their enzyme-bound state. That the agr- and ß-anomeric forms of GlcAc bind to lysozyme in a competitive fashion was indicated by observing the proton magnetic resonance spectra in the presence of 2-acetamido-d3-2-deoxy-agr-d-glucopyranose. The methyl glycosides, methyl-agr-GlcAc and methyl-ß-GlcAc, were also shown to bind competitively with both anomers of GlcAc. Quantitative analysis of the chemical shift data observed for the association of GlcAc with lysozyme was complicated by the mutarotation of GlcAc between its agr- and ß-anomeric forms. However, in the case of the methyl glucosides, where the conformation of each anomer is frozen, it was possible to analyze the chemical shift data in a straightforward manner, and the dissociation constant as well as the chemical shift of the acetamido methyl protons of the enzyme-inhibitor complex was determined for both anomers. The results indicate that the two anomers of methyl-GlcAc bind to lysozyme with slightly different affinities but that the acetamido methyl groups of both anomers experience identical magnetic environments in the enzyme-inhibitor complex

Robustness to Algorithmic Singularities and Sensitivity in Computational Kinematics

A robust approach to computational kinematics intended to cope with algorithmic singularities is introduced in this article. The approach is based on the reduction of the original system of equations to a subsystem of bivariate equations, as opposed to the multivariate polynomial reduction leading to the characteristic univariate polynomial. The effectiveness of the approach is illustrated for the exact function-generation synthesis of planar, spherical, and spatial four-bar linkages. Some numerical examples are provided for the case of the spherical four-bar function generator with six precision points to show the benefits of the proposed method with respect to methods reported in the literature.The first author acknowledges the support of Universidad Politecnica de Valencia, research project PAID-00-09. The second author acknowledges the support of McGill University by means of a James McGill Professorship.Gracia Calandin, LI.; Angeles, J. (2011). Robustness to Algorithmic Singularities and Sensitivity in Computational Kinematics. Proceedings of the Institution of Mechanical Engineers part C - Journal of Mechanical Engineering Science. 225(4):987-999. doi:10.1243/09544062JMES2464S9879992254Raghavan, M., & Roth, B. (1993). Inverse Kinematics of the General 6R Manipulator and Related Linkages. Journal of Mechanical Design, 115(3), 502-508. doi:10.1115/1.2919218Lee, H. Y., Woernle, C., & Hiller, M. (1991). A Complete Solution for the Inverse Kinematic Problem of the General 6R Robot Manipulator. Journal of Mechanical Design, 113(4), 481-486. doi:10.1115/1.2912808Innocenti, C., & Parenti-Castelli, V. (1993). Echelon form solution of direct kinematics for the general fully-parallel spherical wrist. Mechanism and Machine Theory, 28(4), 553-561. doi:10.1016/0094-114x(93)90035-tGosselin, C. M., Sefrioui, J., & Richard, M. J. (1994). On the Direct Kinematics of Spherical Three-Degree-of-Freedom Parallel Manipulators of General Architecture. Journal of Mechanical Design, 116(2), 594-598. doi:10.1115/1.2919419Alizade, R. I., & Kilit, Ö. (2005). Analytical synthesis of function generating spherical four-bar mechanism for the five precision points. Mechanism and Machine Theory, 40(7), 863-878. doi:10.1016/j.mechmachtheory.2004.12.010Cervantes-Sánchez, J. J., Gracia, L., Rico-Martínez, J. M., Medellín-Castillo, H. I., & González-Galván, E. J. (2009). A novel and efficient kinematic synthesis approach of the spherical 4R function generator for five and six precision points. Mechanism and Machine Theory, 44(11), 2020-2037. doi:10.1016/j.mechmachtheory.2009.05.006Angeles, J. (2007). Fundamentals of Robotic Mechanical Systems. Mechanical Engineering Series. doi:10.1007/978-0-387-34580-2Bai, S., Hansen, M. R., & Angeles, J. (2009). A robust forward-displacement analysis of spherical parallel robots. Mechanism and Machine Theory, 44(12), 2204-2216. doi:10.1016/j.mechmachtheory.2009.07.005Sommese, A. J., & Wampler, C. W. (2005). The Numerical Solution of Systems of Polynomials Arising in Engineering and Science. doi:10.1142/5763Forsythe, G. E. (1970). Pitfalls in Computation, or why a Math Book isn’t Enough. The American Mathematical Monthly, 77(9), 931. doi:10.2307/2318109Angeles, J., Hommel, G., & Kovács, P. (Eds.). (1993). Computational Kinematics. Solid Mechanics and Its Applications. doi:10.1007/978-94-015-8192-9Yang, A. T., & Freudenstein, F. (1964). Application of Dual-Number Quaternion Algebra to the Analysis of Spatial Mechanisms. Journal of Applied Mechanics, 31(2), 300-308. doi:10.1115/1.3629601Gupta, K. C., & Beloiu, A. S. (1998). Branch and circuit defect elimination in spherical four-bar linkages. Mechanism and Machine Theory, 33(5), 491-504. doi:10.1016/s0094-114x(97)00078-

The moderating effect of brand orientation on inter-firm market orientation and performance

While prior research has shown that market and brand orientation are key contributors to successful business performance, research to date has not fully explored how inter firm collaboration for these two key orientations can enhance business performance. The purpose of the paper is to investigate the relationship between inter-firm market and performance; to test for the moderating role of brand orientation in that relationship. A total of 169 completed pairs of surveys were collected of small and medium enterprises operating internationally in a variety of industries in Switzerland. The results show that inter-firm market and brand orientation are two antecedents of marketing and financial performance. The impact of inter-firm market on marketing and financial performance is significant when the brand orientation is favorable. This study extends previous research by examining the moderating role of brand orientation on inter firm market orientation, which is important, especially for firms wanting to increase their brand reputation by entering into partnerships with other firms. Further research is indicated, to identify the key moderators of the driving force of inter-firm market in relation to business performance and the reason why maintaining a strong brand presence is important in the international marketplace

Adhesion between elastic solids with randomly rough surfaces: comparison of analytical theory with molecular dynamics simulations

The adhesive contact between elastic solids with randomly rough, self affine fractal surfaces is studied by molecular dynamics (MD) simulations. The interfacial binding energy obtained from the simulations of nominally flat and curved surfaces is compared with the predictions of the contact mechanics theory by Persson. Theoretical and simulation results agree rather well, and most of the differences observed can be attributed to finite size effects and to the long-range nature of the interaction between the atoms in the block and the substrate in the MD model, as compared to the analytical theory which is for an infinite system with interfacial contact interaction. For curved surfaces (JKR-type of problem) the effective interfacial energy exhibit a weak hysteresis which may be due to the influence of local irreversible detachment processes in the vicinity of the opening crack tip during pull-off.Comment: 6 pages, 6 figure

An algebraic/numerical formalism for one-loop multi-leg amplitudes

We present a formalism for the calculation of multi-particle one-loop amplitudes, valid for an arbitrary number N of external legs, and for massive as well as massless particles. A new method for the tensor reduction is suggested which naturally isolates infrared divergences by construction. We prove that for N>4, higher dimensional integrals can be avoided. We derive many useful relations which allow for algebraic simplifications of one-loop amplitudes. We introduce a form factor representation of tensor integrals which contains no inverse Gram determinants by choosing a convenient set of basis integrals. For the evaluation of these basis integrals we propose two methods: An evaluation based on the analytical representation, which is fast and accurate away from exceptional kinematical configurations, and a robust numerical one, based on multi-dimensional contour deformation. The formalism can be implemented straightforwardly into a computer program to calculate next-to-leading order corrections to multi-particle processes in a largely automated way.Comment: 71 pages, 7 figures, formulas for rank 6 pentagons added in Appendix

On a q-extension of Mehta's eigenvectors of the finite Fourier transform for q a root of unity

It is shown that the continuous q-Hermite polynomials for q a root of unity have simple transformation properties with respect to the classical Fourier transform. This result is then used to construct q-extended eigenvectors of the finite Fourier transform in terms of these polynomials.Comment: 12 pages, thoroughly rewritten, the q-extended eigenvectors now N-periodic with q an M-th root of

NMR studies of the phosphotransfer domain of the histidine kinase CheA from Escherichia coli: assignments, secondary structure, general fold, and backbone dynamics

Multidimensional heteronuclear NMR techniques were applied to study the phosphotransfer domain, residues 1 - 134, of the histidine kinase CheA, from Escherichia coli, which contains the site of autophosphorylation, His48. Assignments of the backbone amide groups and side chain patons are nearly complete. Our studies show that this protein fragment consists of five a-helices (A-E)connected by turns. Analysis of NOE distance restraints provided by two-dimensional (2D) ^1H-^1H and three-dimensional (3D) ^(15)N-edited NOESY spectra using model building and structure calculations indicates that the five helices form an antiparallel helix bundle with near-neighbor connectivity. The amino-tenninal four helices are proposed to be arranged in a right-handed manner with helix E packing against helices C and D. From ideal hydrophobic helical packing and structure calculations, the site of autophosphorylation, His48, is nearly fully exposed to the solvent. We measured the NMR relaxation properties of the backbone ^(15)N nuclei using inverse detected two-dimensional NMR spectroscopy. The protein backbone dynamics studies show that CheA1-134 is formed into a tight and compact structure with very limited flexibilities both in helices and turns. Structural implications of titration and phosphorylation experiments are briefly discussed

Phosphotransfer and CheY-Binding Domains of the Histidine Autokinase CheA Are Joined by a Flexible Linker

Multidimensional heteronuclear NMR techniques were applied to study a protein fragment of the histidine autokinase CheA from Escherichia coli. This fragment (CheA_(1-233)) contains the phosphotransfer domain and the CheY-binding domain joined by a linker region. Comparison of chemical shift and NOE cross-peak patterns indicates that the structures of the two domains in CheA_(1-233) remain nearly the same as in the two individual domain fragments, CheA1-134 and CheA_(124-257). Relaxation properties of the backbone ^(15)N nuclei were measured to study the rotational correlations of the two domains and properties of the linker region. Dynamics data were analyzed both by an isotropic motional model and an anisotropic motional model. The experimental T_1 and T_2 values, the derived rotational correlation times, and motional anisotropy are significantly different for the two domains, indicating the two domains reorient independently and the linker region is highly flexible. Dynamics data of CheA_(1-233) were also compared with those of CheA_(1-134). Our studies show that flexible domain linkers and extended and flexible terminal polypeptide chains can have significant effects on the motional properties of the adjacent structured regions. These observations suggest a model for the graded regulation of CheA autophosphorylation activity. In this model, the various activity states of the receptor are generated by controlling the access of the mean position of the kinase domain to the phosphotransfer domain. This would then modulate the diffusional encounter rate of the domains and hence activity over a wide and graded range of values