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31.
Solution NMR approaches for establishing specificity of weak heterodimerization of membrane proteins
Solution NMR provides a powerful approach for detecting complex formation involving weak to moderate intermolecular affinity. However, solution NMR has only rarely been used to detect complex formation between two membrane proteins in model membranes. The impact of specific binding on the NMR spectrum of a membrane protein can be difficult to distinguish from spectral changes that are induced by nonspecific binding and/or by changes that arise from forced cohabitation of the two proteins in a single model membrane assembly. This is particularly the case when solubility limits make it impossible to complete a titration to the point of near saturation of complex formation. In this work experiments are presented that provide the basis for establishing whether specific complex formation occurs between two membrane proteins under conditions where binding is not of high avidity. Application of these methods led to the conclusion that the membrane protein CD147 (also known as EMMPRIN or basigin) forms a specific heterodimeric complex in the membrane with the 99-residue transmembrane C-terminal fragment of the amyloid precursor protein (C99 or APP-βCTF), the latter being the immediate precursor of the amyloid-β polypeptides that are closely linked to the etiology of Alzheimer's disease. 相似文献
32.
We describe the use of dynamic combinatorial chemistry to discover a new series of linear hydrazone-based receptors that bind multiple dihydrogen phosphate ions. Through the use of a template-driven, selection-based approach to receptor synthesis, dynamic combinatorial chemistry allows for the identification of unexpected host structures and binding motifs. Notably, we observed the unprecedented selection of these linear receptors in preference to competing macrocyclic hosts. Furthermore, linear receptors containing up to nine building blocks and three different building blocks were amplified in the dynamic combinatorial library. The receptors were formed using a dihydrazide building block based on an amino acid-disubstituted ferrocene scaffold. A detailed study of the linear pentamer revealed that it forms a helical ditopic receptor that employs four acylhydrazone hydrogen-bond donor motifs to cooperatively bind two dihydrogen phosphate ions. 相似文献
33.
B. J. van der Veken R. S. Sanders J. Xiao J. R. Durig 《Journal of Molecular Structure》2001,560(1-3):57-86
Infrared spectra (4000–50 cm−1) of the vapor, amorphous and crystalline solids and Raman spectra (3600–10 cm−1) of the liquid with qualitative depolarization data as well as the amorphous and crystalline solids of methylaminothiophosphoryl difluoride, CH3N(H)P(=S)F2, and three deuterated species, CD3N(H)P(=S)F2, CH3N(D)P(=S)F2, and CD3N(D)P(=S)F2, have been recorded. The spectra indicate that in the vapor, liquid and amorphous solid a small amount of a second conformer is present, whereas only one conformer remains in the low temperature crystalline phase. The near-infrared spectra of the vapor confirms the existence of two conformers in the gas phase. Asymmetric top contour simulation of the vapor shows that the trans conformer is the predominant vapor phase conformer. From a temperature study of the Raman spectrum of the liquid the enthalpy difference between the trans and near-cis conformers was determined to be 368±15 cm−1 (4.41±0.2 kJ/mol), with the trans conformer being thermodynamically preferred. Ab Initio calculations with structure optimization using the 6-31G(d) and 6-311+G(d,p) basis sets at the restricted Hartree–Fock (RHF) and/or with full electron correlation by the perturbation method to second order (MP2) support the occurrence of near-trans (5° from trans) and near-cis (20° from cis) conformers. From the RHF/6-31G(d) calculation the near-trans conformer is predicted to be the more stable form by 451 cm−1 (5.35 kJ/mol) and from the MP2/6-311+G(d,p) calculation by 387 cm−1 (4.63 kJ/mol). All of the normal modes of the near-trans rotamer have been assigned based on infrared band contours, depolarization values and group frequencies and the assignment is supported by the normal coordinate calculation utilizing harmonic force constants from the MP2/6-31G(d) ab initio calculations. 相似文献
34.
35.
A cyclic porphyrin tetramer, consisting of two bis-phosphine substituted zinc(II) porphyrin units and two Rh(III)TPP units, is selected and amplified virtually quantitatively from a dynamic combinatorial library using 4,4'-bipy as a scaffold and using orthogonal binding modes. 相似文献
36.
Gorzelle BM Hoffman AK Keyes MH Gray DN Ray DG Sanders CR 《Journal of the American Chemical Society》2002,124(39):11594-11595
Amphipathic polymers ("amphipols") were introduced several years ago (Tribet, C.; Audebert, R.; Popot, J.-L. Proc. Natl. Acad. Sci. U.S.A. 1996, 93, 15047-15050) as an alternative method for solubilizing integral membrane proteins in stable, nativelike conformations. However, direct maintenance of full membrane protein functionality in amphipol solutions has not previously been demonstrated in the absence of added lipid or detergent. In this contribution, the first zwitterionic amphipol "PMAL-B-100" is introduced. PMAL-B-100 not only maintains membrane protein structure and solubility, but also supports the full catalytic activity of an integral membrane enzyme, diacylglycerol kinase, in the complete absence of additional lipid or detergent. All of the roles which a lipid bilayer normally plays in maintaining diacylglycerol kinase's structure and in facilitating catalysis are satisfied by the environment and interactions supplied by PMAL-B-100. 相似文献
37.
The clinically effective anticancer agent hexamethylmelamine undergoes an unusual acylhydroperoxide oxidative transformation to give the novel product 2,4-bis(dimethylamino)-6-[(dimethylamino)oxy]-1,3,5-triazine. 相似文献
38.
The first clear evidence for the existence of from a high resolution, far-infrared (FIR) study of the antiferromagnetic resonance (AFMR) of FeF2 is presented. The broad (4 kOe) asymmetric structure observed in transmission is shown to be a characteristic polariton effect; the linewidth ΔH at 4.2 K is only 350 Oe. The Mn impurity mode is enhanced by ~50 in agreement with a coupled equations of motion prediction. Both host and impurity resonances are broadened by superradiance. 相似文献
39.
A two-atom blocking model has been developed to analyze surface blocking patterns recently obtained in medium energy ion scattering experiments. In the model the mean square thermal vibration amplitude of a surface atom can be used as a fitting parameter in comparison with experimental results. Anisotropic surface vibrations have been taken into account by assuming a non-spherical Gaussian probability distribution for the thermal displacement of the surface atom. In addition, some analytical expressions have been derived for the effective nuber of target atoms contributing to the yield of ions backscattered into a blocking direction. 相似文献
40.