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71.
D. L. Turner Lorraine Brennan Stephen G. Chamberlin Ricardo O. Louro António V. Xavier 《European biophysics journal : EBJ》1998,27(4):367-375
Standard procedures for using nuclear Overhauser enhancements (NOE) between protons to generate structures for diamagnetic
proteins in solution from NMR data may be supplemented by using dipolar shifts if the protein is paramagnetic. This is advantageous
since the electron-nuclear dipolar coupling provides relatively long-range geometric information with respect to the paramagnetic
centre which complements the short-range distance constraints from NOEs. Several different strategies have been developed
to date, but none of these attempts to combine data from NOEs and dipolar shifts in the initial stages of structure calculation
or to determine three dimensional protein structures together with their magnetic properties. This work shows that the magnetic
and atomic structures are highly correlated and that it is important to have additional constraints both to provide starting
parameters for the magnetic properties and to improve the definition of the best fit. Useful parameters can be obtained for
haem proteins from Fermi contact shifts; this approach is compared with a new method based on the analysis of dipolar shifts
in haem methyl groups with respect to data from horse and tuna ferricytochromes c. The methods developed for using data from NOEs and dipolar shifts have been incorporated in a new computer program, PARADYANA,
which is demonstrated in application to a model data set for the sequence of the haem octapeptide known as microperoxidase-8.
Received: 13 October 1997 / Accepted: 19 December 1997 相似文献
72.
Lucia Banci I. Bertini J. Gaspard Huber Georgios A. Spyroulias Paola Turano 《Journal of biological inorganic chemistry》1999,4(1):21-31
In the frame of a broad study on the structural differences between the two redox forms of cytochromes to be related to the
electron transfer process, the NMR solution structure of horse heart cytochrome c in the reduced form has been determined. The structural data obtained in the present work are compared to those already available
in the literature on the same protein and the presence of conformational differences is discussed in the light of the experimental
method employed for the structure determination. Redox-state dependent changes are analyzed and in particular they are related
to the role of propionate-7 of the heme. Also some hydrogen bonds are changed upon reduction of the heme iron. A substantial
similarity is observed for the backbone fold, independently of the oxidation state. At variance, some meaningful differences
are observed in the orientation of a few side chains. These changes are related to those found in the case of the highly homologous
cytochrome c from Saccharomyces cerevisiae. The exchangeability of the NH protons has been investigated and found to be smaller than in the case of the oxidized protein.
We think that this is a characteristic of reduced cytochromes and that mobility is a medium for molecular recognition in vivo.
Received: 8 June 1998 / Accepted: 13 October 1998 相似文献
73.
Mengli Cai Ying Huang Jianhua Liu Ramaswamy Krishnamoorthi 《Journal of biomolecular NMR》1995,6(2):123-128
Summary Three different conformations of proline rings in a protein in solution, Up, Down and Twist, have been distinguished, and stereospecific assignments of the pyrrolidine -, - and -hydrogens have been made on the basis of 1H-1H vicinal coupling constant patterns and intraresidue NOEs. For all three conformations, interhydrogen distances in the pairs -3, 3-3, 2-2, 2-2, and 3-3 (2.3 Å) are shorter than those in the pairs -2, 2-3, 3-2, 2-3, and 3-2 (2.7–3.0 Å), resulting in stronger NOESY cross peaks. For the Up conformation, the 3-2 and 2-3 spin-spin coupling constants are small (<3 Hz), and weak cross peaks are obtained in a short-mixing-time (10 ms) TOCSY spectrum; all other vicinal coupling constants are in the range 5–12 Hz, and result in medium to strong TOCSY cross peaks. For the Down form, the -2, 2-3, and 3-2 vicinal coupling constants are small, leading to weak TOCSY cross peaks; all other couplings again are in the range 5–12 Hz, and result in medium to strong TOCSY cross peaks. In the case of a Twist conformation, dynamically averaged coupling constants are anticipated. The procedure has been applied to bovine pancreatic trypsin inhibitor and Cucurbita maxima trypsin inhibitor-V, and ring conformations of all prolines in the two proteins have been determined. 相似文献
74.
75.
Summary The solution conformation of human big endothelin-1, a 38-residue peptide which serves as the putative precursor to the potent vasoconstrictor endothelin-1 has been examined by1H NMR. NOEs were utilized as distance restraints in the distance geometry program DSPACE to generate initial structures. Further refinement of these structures was accomplished through molecular mechanics/molecular dynamics in an iterative process involving the incorporation of stereospecific assignments of prochiral centers and the use of back-calculation of NOESY spectra. A family of structures consisting of a type 11 -turn for residues 5–8 and an -helix extending from residues 9–16 constitute a well-defined region, as reflected by the atomic root-mean-square (RMS) difference of 1.56 Å about the mean coordinate positions of the backbone atoms (N, C, Ca and O). This core region (residues 1-15) is very similar to the core residues of endothelin-1 (Donlan, M. et al. (1991)J. Cell. Biochemistry, S15G, 85). While the evidence from NOESY and coupling constant data suggests that the C-terminal region, residues 17–34, is not a mixture of randomly distributed chain conformations, it is also not consistent with a single chain conformation. Under the conditions studied, residues 17–38 in human big endothelin-1 in water at pH 3.0 between 20–30°C appear to be represented by a series of conformers in dynamic equilibrium. 相似文献
76.
Giastas P Eliadou K Plyta ZF Yannakopoulou K Mavridis IM 《Carbohydrate research》2004,339(6):1189-1194
Heptakis(2,3-di-O-acetyl-6-bromo-6-deoxy)cyclomaltoheptaose has been characterized in aqueous solution by 1D and 2D NMR spectroscopy and in the solid state by X-ray crystallography. In methanol solution, the acetyl groups were found to interact with both inward and outward-pointing protons. This and the strong deshielding of the bridging carbons, relative to the nonacetylated precursor, indicate macrocyclic flexibility. In the crystalline state the macrocycle exists as a methanol complex. It exhibits elliptical distortion, all glucose residues been tilted with their primary side toward the cavity. The existing strain due to the congestion of 14 acetyl groups at the secondary site is relieved by two glucose rings acquiring the rarely observed skew-boat conformation, (0)S(2), by the increased tilting of two glucose residues, as well as by minor variations of the torsion angles of the acetyl groups. The seven bromine atoms are quite accessible to nucleophiles. 相似文献
77.
电渗析技术用于猪毛水解液脱酸的研究 总被引:3,自引:1,他引:3
利用上海华凯科技贸易公司制作的HF系列离子交换膜为隔膜的电渗析器对猪毛水解液进行电渗析脱酸,研究了脱酸过程中浓缩室、淡化室酸度变化及电流效率 相似文献
78.
Aneta Czy? Jan Fronk Krzysztof Zab?ocki 《Biochemical and biophysical research communications》2009,381(2):176-2564
Tunicamycin effect on thapsigargin-induced store-operated calcium entry was investigated. Ca2+ influx was stimulated by 50% upon exposure of Jurkat cells to tunicamycin. Moreover, tunicamycin efficiently prevented the inhibition of store-operated calcium entry caused by dissipation of mitochondrial membrane potential. Protective action of tunicamycin on store-operated Ca2+ entry was also partially preserved in Jurkat cells depleted of ATP, while Ca2+ entry into ATP-deprived cells grown in tunicamycin-free medium was almost completely inhibited. Tunicamycin-evoked changes in cellular Ca2+ fluxes coincided with decreased glycosylation of STIM1 protein. Although the latter observation is correlative and needs additional confirmation it may suggest that deglycosylation of STIM1 protein deprives store-operated calcium entry system of an important regulatory mechanism. This study suggests a novel mechanism of modulation of the activity of store-operated calcium channels in lymphoidal cells. 相似文献
79.
Complexation of desferrioxamine B (DFB) and desferricoprogen (DFC) with Cd(II) and Pb(II) toxic ions as well as complexation of DFC with Ca(II) and Mg(II) essential metals have been investigated and the results have been compared to those with other metal ions. The two siderophores have moderate Cd(II)-binding ability, but both, and especially DFC, bind Pb(II) in high stability complexes. Surprisingly, significant differences exist between Pb(II)-complexation of DFB and DFC. Namely, a maximum of two hydroxamate groups of a DFB coordinate to a Pb(II) ion, the third one binds to another metal ion with high preference and the formation of a trinuclear species, [Pb3(DFBH)2]2+, is predominant even at 1:1 metal to ligand ratio in this system. On the contrary, DFC forms mononuclear complex, [ML], with much higher stability and the formation of the trinuclear complex is negligible compared to DFB. The 6s2 electron-pair of Pb(II), which remains always inert during complexation with hydroxamic acids and also with DFB, seems to become active in the DFC complexes (due to the effect of the double bonds in β-position to each hydroxamate), what, at least in some extent, allows the coordination of all the three hydroxamates of DFC to the same Pb(II) ion. This way of interaction (unique with a hydroxamate-based compound) results in significant stability increase, and, as a consequence, DFC is much better Pb(II)-chelating agent than DFB. Although DFC forms unexpectedly high stability complexes with Mg(II) compared to Ca(II), but even Mg(II), compared to many other metals, is not an efficient DFC-binding metal. Therefore, any sequestration of this biologically very important metal is not likely from a living organism by DFC. 相似文献
80.
Mateja Zorko 《生物化学与生物物理学报:生物膜》2009,1788(2):314-21276
We have produced a small antimicrobial peptide PFWRIRIRR in bacteria utilizing production in the form of insoluble fusion protein with ketosteroid isomerase. The recombinant peptide was rapidly and efficiently isolated by acidic cleavage of the fusion protein based on the acid labile Asp-Pro bond at the N-terminus of the peptide. The peptide has antibacterial activity and neutralizes macrophage activation by LPS. The selectivity of the peptide against bacteria correlates with preferential binding to acidic phospholipid vesicles. Solution structure of the peptide in SDS and DPC micelles was determined by NMR. The peptide adopts a well-defined structure, comprising a short helical segment. Cationic and hydrophobic clusters are segregated along the molecular axis of the short helix, which is positioned perpendicular to the membrane plane. The position of the helix is shifted in two micellar types and more nonpolar surface is exposed in anionic micelles. Overall structure explains the advantageous role of the N-terminal proline residue, which forms an integral part of the hydrophobic cluster. 相似文献