共查询到20条相似文献,搜索用时 156 毫秒
1.
Youxing Qu Songping Liang Jizhen Ding Libin Ma Renji Zhang Xiaocheng Gu 《The protein journal》1995,14(7):549-557
The complete sequence-specific assignments of resonances in the1H-NMR spectrum of huwentoxin-I from the Chinese bird spider,Selenocosmia huwena, is described. A combination of two-dimensional NMR experiments including 2D-COSY, 2D-NOESY, and 2D-TOCSY has been employed on samples of the toxin dissolved in D2O and in H2O for assignment purposes. Protons belonging to spin systems for each of the 33 amino acids were identified. The sequence-specific assignments were facilitated by the identification ofd αN connectivities on the fingerprint regions of the COSY and NOESY spectra and were supported by the identification ofd NN andd αN connectivities in the TOCSY and NOESY spectra. These studies provide a basis for the determination of the solution-phase conformation of this toxin. 相似文献
2.
Summary Application of 1H 2D NMR methods to solubilized membrane proteins and peptides has up to now required the use of selectively deuterated detergents. The unavailability of any of the common biochemical detergents in deuterated form has therefore limited to some extent the scope of this approach. Here a 1H NMR method is described which allows structure determination of membrane peptides and small membrane proteins by 1H 2D NMR in any type of non-deuterated detergent. The approach is based on regioselective excitation of protein resonances with DANTE-Z or spin-pinging pulse trains. It is shown that regioselective excitation of the amide-aromatic region of solubilized membrane proteins and peptides leads to an almost complete suppression of the two orders of magnitude higher contribution of the protonated detergent to the 1H NMR spectrum. Consistently TOCSY, COSY and NOESY sequences incorporating such regioselective excitation in the F2 dimension yield protein 1H 2D NMR spectra of quality comparable to those obtained in deuterated detergents. Regioselective TOCSY and NOESY spectra display all through-bond and through-space correlations within amide-aromatic protons and between these protons and aliphatic and -protons. Regioselective COSY spectra provide scalar coupling constants between amide and -protons. Application of the method to the membrane-active peptide mastoparan X, solubilized in n-octylglucoside, yields complete sequence-specific assignments and extensive secondary structure-related spatial proximities and coupling constants. It is shown that mastoparan adopts an -helical conformation when bound to nonionic detergent micelles. The present method is expected to increase the applicability of 1H solution NMR methods to membrane proteins and peptides.Abbreviations 2D NMR
two-dimensional NMR
- COSY
correlated spectroscopy
- DANTE
delays alternating nutations for tailored excitation
- NOESY
nuclear Overhauser enhancement spectroscopy
- TOCSY
total correlation spectroscopy 相似文献
3.
Rasmus H. Fogh Dick Schipper Rolf Boelens Robert Kaptein 《Journal of biomolecular NMR》1995,5(3):259-270
Summary The 1H, 13C and 15N NMR resonances of serine protease PB92 have been assigned using 3D tripleresonance NMR techniques. With a molecular weight of 27 kDa (269 residues) this protein is one of the largest monomeric proteins assigned so far. The side-chain assignments were based mainly on 3D H(C)CH and 3D (H)CCH COSY and TOCSY experiments. The set of assignments encompasses all backbone carbonyl and CHn carbons, all amide (NH and NH2) nitrogens and 99.2% of the amide and CHn protons. The secondary structure and general topology appear to be identical to those found in the crystal structure of serine protease PB92 [Van der Laan et al. (1992) Protein Eng., 5, 405–411], as judged by chemical shift deviations from random coil values, NH exchange data and analysis of NOEs between backbone NH groups.Abbreviations 2D/3D/4D
two-/three-/four-dimensional
- HSQC
heteronuclear single-quantum coherence
- HMQC
heteronuclear multiple-quantum coherence
- COSY
correlation spectroscopy
- TOCSY
total correlation spectroscopy
- NOE
nuclear Overhauser enhancement (connectivity)
- NOESY
2D NOE spectroscopy
Experiment nomenclature (H(C)CH, etc.) follows the conventions used elsewhere [e.g. Ikura et al. (1990) Biochemistry, 29, 4659–4667]. 相似文献
4.
K. D. Hahn F. Buck H. Rüterjans B. K. Chernov K. G. Skryabin M. P. Kirpichnikov 《European biophysics journal : EBJ》1985,12(2):87-95
The 17 base pair operator O
R
3 oligonucleotide, which is the preferential binding site for the Cro repressor of phage , was studied by two-dimensional NMR spectroscopy. A sequential assignment procedure based on two-dimensional Nuclear Overhauser Effect (NOESY) and scalar coupling correlated (COSY) NMR spectroscopy, together with the knowledge of the oligodesoxynucleotide sequence, made it possible to assign the non-exhangeable base protons and the H1 and the H2-H2 sugar protons of the O
R
3 operator DNA. The pattern of the observed NOE connectivities is consistent with a right-handed helical DNA structure. The base and sugar proton assignments provide the necessary information for further studies of the O
R
3 operator — Cro repressor interaction.Abbreviations COSY
correlated spectroscopy
- FID
free induction decay
- NOE
nuclear Overhauser effect
- NOESY
nuclear Overhauser effect spectroscopy
- RD
relaxation delay
- TSP
sodium 3-trimethylsilyl-(2,2,3,3-2H4)propionate
- EDTA
sodium ethylendiamine tetraacetate 相似文献
5.
Kerstin Nordstrand Hannes Ponstingl Arne Holmgren Gottfried Otting 《European biophysics journal : EBJ》1996,24(3):179-184
Virtually complete sequence specific 1H and 15N resonance assignments are presented for acid denatured reduced E. coli glutaredoxin 3. The sequential resonance assignments of the backbone rely on the combined use of 3D F1-decoupled ROESY-15N-HSQC and 3D 15N-HSQC-(TOCSY-NOESY)-15N-HSQC using a single uniformly 15N labelled protein sample. The sidechain resonances were assigned from a 3D TOCSY-15N-HSQC and a homonouclear TOCSY spectrum. The presented assignment strategy works in the absence of chemical exchange peaks with signals from the native conformation and without 13C/15N double labelling. Chemical shifts, 3J(H, NH) coupling constants and NOEs indicate extensive conformational averaging of both backbone and side chains in agreement with a random coil conformation. The only secondary structure element persisting at pH 3.5 appears to be a short helical segment comprising residues 37 to 40.Abbreviations HSQC
heteronuclear single quantum coherence
- NMR
nuclear magnetic resonance
- NOE
nuclear Overhauser effect
- NOESY
two-dimensional NOE spectroscopy
- ROE
nuclear Overhauser effect in the rotating frame
- ROESY
two-dimensional ROE spectroscopy
- TOCSY
total correlation spectroscopy
- TPPI
time proportional phase incrementation
Correspondence to: G. Otting 相似文献
6.
Ivano Bertini Bertini Luchinat Mario Piccioli Margarita Vicens Oliver Maria Silvia Viezzoli 《European biophysics journal : EBJ》1991,20(5):269-279
Human copper-cobalt superoxide dismutase in the reduced form has been investigated through 1H NMR techniques. The aim is to monitor the structural properties of this derivative and to compare them with those of reduced and oxidized native superoxide dismutases. The observed signals of the cobalt ligands have been assigned as well as the signals of the histidines bound to copper(I). The latter signals experience little pseudocontact shifts which allow a rough orientation of the magnetic susceptibility tensor in the molecular frame. The connectivities indicate that, although the histidine bridge is broken in the reduced form, the interproton distances between ligands of both ions are essentially the same.Abbreviations WEFT
water eliminated Fourier transform
- NOE
nuclear Overhauser effect
- NOESY
NOE spectroscopy
- COSY
correlation spectroscopy
- TOCSY
total correlation spectroscopy
- SOD
superoxide dismutase
- E2Co(II)SOD
SOD with empty copper site (E=empty) and with cobalt(II) in the Zinc(II) site
Offprint requests to: I. Bertini 相似文献
7.
The program XEASY for computer-supported NMR spectral analysis of biological macromolecules 总被引:16,自引:0,他引:16
Summary A new program package, XEASY, was written for interactive computer support of the analysis of NMR spectra for three-dimensional structure determination of biological macromolecules. XEASY was developed for work with 2D, 3D and 4D NMR data sets. It includes all the functions performed by the precursor program EASY, which was designed for the analysis of 2D NMR spectra, i.e., peak picking and support of sequence-specific resonance assignments, cross-peak assignments, cross-peak integration and rate constant determination for dynamic processes. Since the program utilizes the X-window system and the Motif widget set, it is portable on a wide range of UNIX workstations. The design objective was to provide maximal computer support for the analysis of spectra, while providing the user with complete control over the final resonance assignments. Technically important features of XEASY are the use and flexible visual display of strips, i.e., two-dimensional spectral regions that contain the relevant parts of 3D or 4D NMR spectra, automated sorting routines to narrow down the selection of strips that need to be interactively considered in a particular assignment step, a protocol of resonance assignments that can be used for reliable bookkeeping, independent of the assignment strategy used, and capabilities for proper treatment of spectral folding and efficient transfer of resonance assignments between spectra of different types and different dimensionality, including projected, reduced-dimensionality triple-resonance experiments.Abbreviations 1D, 2D, 3D, 4D
one-, two-, three-, four-dimensional
- NOE
nuclear Overhauser enhancement
- NOESY
nuclear Overhauser enhancement spectroscopy
- TOCSY
total correlation spectroscopy
- COSY
correlation spectroscopy
- TPPI
time-proportional phase incrementation 相似文献
8.
Chinpan Chen Kellie Hom Rong-Fong Huang Ping-Jung Chou You-Di Liao Tai-huang Huang 《Journal of biomolecular NMR》1996,8(3):331-344
Summary RC-RNase is a pyrimidine-guanine sequence-specific ribonuclease and a sialic-acid-binding lectin purified from Rana catesbeiana (bullfrog) oocytes. This 111-amino acid protein exhibits cytotoxicity toward several tumor cell lines. In this paper we report the assignments of proton NMR resonances and the identification of the secondary structure deduced from NOE constraints, chemical shift index, 3JNH and amide proton exchange rates. The protein was directly isolated from bullfrog oocytes; we were able to assign all but five of the amino acid backbone protons of the unlabeled protein by analyzing a large set of two-dimensional proton NMR spectra obtained at several temperatures and pH conditions. Our results indicate that the structure of RC-RNase is dominated by the presence of two triple-stranded antiparallel -sheets and three -helices, similar to those of the pyrimidine family ribonucleases. Two sets of resonances were observed for 11 amide protons and 8 -protons located in the loop-1 region, an 2 helix, and three -strands (1, 3 and 4), suggesting the presence of nonlocalized multiple conformations for RC-RNase.Abbreviations DQF-COSY
double-quantum-filtered correlation spectroscopy
- DTT
dithiothreitol
- NOE
nuclear Overhauser enhancement
- NOESY
nuclear Overhauser enhancement spectroscopy
- PE-1
N-terminal pyroglutamate
- RC-RNase
ribonuclease from the oocyte of Rana catesbeiana
- TOCSY
total correlation spectroscopy
- TPPI
time-proportional phase incrementation
- TSP
sodium 3-trimethylsilylpropionate-2,2,3,3-d
4 相似文献
9.
Summary Sequence-specific 1H and 15N resonance assignments have been made for 137 of the 146 nonprolyl residues in oxidized Desulfovibrio desulfuricans [Essex 6] flavodoxin. Assignments were obtained by a concerted analysis of the heteronuclear three-dimensional 1H-15N NOESY-HMQC and TOCSY-HMQC data sets, recorded on uniformly 15N-enriched protein at 300 K. Numerous side-chain resonances have been partially or fully assigned. Residues with overlapping 1HN chemical shifts were resolved by a three-dimensional 1H-15N HMQC-NOESY-HMQC spectrum. Medium-and long-range NOEs, 3JNH
coupling constants, and 1HN exchange data indicate a secondary structure consisting of five parallel -strands and four -helices with a topology similar to that of Desulfovibrio vulgaris [Hidenborough] flavodoxin. Prolines at positions 106 and 134, which are not conserved in D. vulgaris flavodoxin, contort the two C-terminal -helices.Abbreviations CSI
chemical shift index
- DQF-COSY
double-quantum-filtered correlation spectroscopy
- DIPSI
decoupling in the presence of scalar interactions
- FMN
flavin mononucleotide
- GARP
globally optimized alternating phase rectangular pulse
- HMQC
heteronuclear multiple-quantum coherence
- HSQC
heteronuclear single-quantum coherence
- NOE
nuclear Overhauser effect
- NOESY
nuclear Overhauser enhancement spectroscopy
- TOCSY
total correlation spectroscopy
- TPPI
time-proportional phase increments
- TSP
3-(trimethylsilyl)propionic-2,2,3,3-d
4 acid, sodium salt 相似文献
10.
Two-dimensional 1H-NMR investigation of the water conformation of the atrial natriuretic factor (ANF 101-126) 总被引:1,自引:0,他引:1
The complete sequential assignment of the 1H-nmr resonance frequencies of the active fragment of the rat atrial natriuretic factor (ANF 101–126) has been performed. Two-dimensional nmr techniques have been employed, including phase-sensitive nuclear Overhauser spectroscopy (NOESY), relayed coherence transfer spectroscopy (RELAY), and J-correlated spectroscopy (COSY). Experiments were performed both in D20 and H20 solutions at different pH values. With few exceptions, resonance frequencies were practically pH independent. NOESY spectra were recorded using both 300- and 500-ms mixing times, and no long-range connectivities were observed, leading to the conclusion that ANF 101–126 has no defined secondary nor tertiary structure in water in the pH range used (2.73–5.21). 相似文献
11.
P. Berthault N. Birlirakis G. Rubinstenn P. Sinaÿ H. Desvaux 《Journal of biomolecular NMR》1996,8(1):23-35
Summary A recent 1H NMR method has been applied to the determination of the solution structure and internal dynamics of a synthetic mixed C/O trisaccharide related to sialyl Lewisx. Varying the rf field offset in ROESY-type experiments enabled the measurement of longitudinal and transverse dipolar cross-relaxation rates with high accuracy. Assuming that for each proton pair the motion could be represented by a single exponential autocorrelation function, it was possible to derive geometrical parameters (r) and dynamic parameters cp. With this assumption, 224 cross-relaxation rates have been transformed into 30 interproton distance constraints and 30 dipolar correlation times. The distance constraints have been used in a simulated-annealing procedure. This trisaccharide exhibits a structure close to the O-glycosidic analogue, but its flexibility seems highly reduced. On the basis of the determined structure and dynamics, it is shown that no conformational exchange occurs, the molecule existing in the form of a unique family in aqueous solution. In order to assess the quality of the resulting structures and to validate this new experimental procedure of distance extraction, we finally compare these solution structures to the ones obtained using three different sets of distances deduced from three choices of internal reference. It appears that this procedure allows the determination of the most precise and accurate solution.Abbreviations COSY
correlation spectroscopy
- NOE
nuclear Overhauser enhancement
- NOESY
nuclear Overhauser enhancement spectroscopy; rmsd, root-mean-square deviation
- ROESY
rotating frame Overhauser enhancement spectroscopy
- SLex
sialyl Lewisx
- TOCSY
total correlation spectroscopy 相似文献
12.
Antonio Donaire Zhi-Hao Zhou Michael W. W. Adams Gerd N. La Mar 《Journal of biomolecular NMR》1996,7(1):35-47
Summary The solution molecular structure of the four-iron ferredoxin (Fd) from the hyperthermophilic archaeon Thermococcus litoralis (Tl) has been investigated by 1H NMR spectroscopy. TOCSY and NOESY experiments in H2O, tailored to detect both weakly and strongly relaxed resonances, together with steady-state NOEs in both H2O and D2O, allowed the identification of 58 of the 59 residues, with one residue near the paramagnetic center undetected. It is shown that the contact shifted and strongly relaxed signals for all four cysteines ligated to the paramagnetic cluster can be assigned by standard backbone connectivities that do not require any assumptions about the tertiary structure. Secondary structural elements identified in Tl Fd are a three-stranded antiparallel -strand involving the termini of the protein, a double -strand (also antiparallel), two -helices and four turns. The existence of a disulfide bridge between the nonligated cysteines is also proposed. Dipolar contacts observed in the NOESY maps and by steady-state NOEs between the ligated cysteines and the diamagnetic protein matrix indicate that the overall folding pattern of Tl Fd is very similar to that of the 3Fe ferredoxin from the mesophilic bacterium Desulfovibrio gigas [Kissinger et al. (1991) J. Mol. Biol., 219, 693–723]. The influence of the paramagnetism of the cluster on the relaxation properties of the proton signals of nonligated residues near the cluster, as well as on the ligated cysteines, correlates well with the proximity to the cluster iron(s), as predicted from the crystal structures for homologous protons of other single-cluster ferredoxins. Finally, the potential role of the various identified structural factors in contributing to the hyperthermostability of this protein is discussed.Abbreviations Fd
ferredoxin
- HiPiP
high-potential iron-sulfur proteins
-
Dg
Desulfovibrio gigas
-
Av
Azotobacter vinelandii
-
Pf
Pyrococcus furiosus
-
Tl
Thermococcus litoralis
-
Pa
Peptostreptococcus asaccharolyticus
-
Bt
Bacillus thermoproteolyticus
-
Cp
Clostridium pasteurianum
-
Ca
Clostridium acidi urici
-
Da
Desulfovibrio africanus
-
Tm
Thermatoga maritima
- NOE
nuclear Overhauser effect
- NOESY
2D NOE spectroscopy
- MCOSY
2D magnitude correlation spectroscopy
- TOCSY
total correlation spectroscopy
To whom correspondence should be addressed. 相似文献
13.
14.
Sequence-specific 1H NMR assignments are presented for a non-selfcomplementary 23-base-pair DNA duplex of molecular weight 15,000 daltons, containing the OR3 repressor binding site of bacteriophage as the central core. The NMR techniques used were mainly phase-sensitive two-dimensional NOE and 2Q spectroscopy, the latter to overcome overlap problems within the spectral region of the deoxyribose spin-systems. Direct sequential NOE connectivities are observed between adenine 2H and deoxyribose 1 protons. We propose the use of these connectivities as a check of the assignments of C1 and A2 protons, which have independently been derived via other assignment pathways.Abbreviation COSY
2-dimensional correlated spectroscopy
- NOESY
2-dimensional nuclear-Overhauser-enhancement spectroscopy
- RELAYED-COSY
2-dimensional relayed coherence transfer spectroscopy
- 2Q
two-quantum
- ppm
parts per million
- 23-bp DNA
d-(ATCTATCACCGCAAGGGATAAAT) · d-(ATTTATCCCTTGCGGTGATAGAT)
- 17-bp DNA (OR3)
d-(TATCACCGCAAGGGATA) · d-(TATCCCTTGCGGTGATA)
- di(X;Y)
intra-residue distance between protons X and Y
- ds(X;Y)
sequential distance between protons X and Y located in sequentially neighbouring nucleotides, where the direction from X to Y is always from 5 to 3 相似文献
15.
Siddhartha Sarma Russell J. DiGate Debra L. Banville R. D. Guiles 《Journal of biomolecular NMR》1996,8(2):171-183
Summary Modern multidimensional double- and triple-resonance NMR methods have been applied to assign the backbone and side-chain 13C resonances for both equilibrium conformers of the paramagnetic form of rat liver microsomal cytochrome b
5. The assignment of backbone 13C resonances was used to confirm previous 1H and 15N resonance assignments [Guiles, R.D. et al. (1993) Biochemistry, 32, 8329–8340]. On the basis of short- and medium-range NOEs and backbone 13C chemical shifts, the solution secondary structure of rat cytochrome b
5 has been determined. The striking similarity of backbone 13C resonances for both equilibrium forms strongly suggests that the secondary structures of the two isomers are virtually identical. It has been found that the 13C chemical shifts of both backbone and side-chain atoms are relatively insensitive to paramagnetic effects. The reliability of such methods in anisotropic paramagnetic systems, where large pseudocontact shifts can be observed, is evaluated through calculations of the magnitude of such shifts.Abbreviations DANTE
delays alternating with nutation for tailored excitation
- DEAE
diethylaminoethyl
- DQF-COSY
2D double-quantum-filtered correlation spectroscopy
- EDTA
ethylenediaminetetraacetic acid
- HCCH-TOCSY
3D proton-correlated carbon TOCSY experiment
- HMQC
2D heteronuclear multiple-quantum correlation spectroscopy
- HNCA
3D triple-resonance experiment correlating amide protons, amide nitrogens and alpha carbons
- HNCO
3D triple-resonance experiment correlating amide protons, amide nitrogens and carbonyl carbons
- HNCOCA
3D triple-resonance experiment correlating amide protons, amide nitrogens and alpha carbons via carbonyl carbons
- HOHAHA
2D homonuclear Hartmann-Hahn spectroscopy
- HOHAHA-HMQC
3D HOHAHA relayed HMQC
- HSQC
2D heteronuclear single-quantum correlation spectroscopy
- IPTG
isopropyl thiogalactoside
- NOESY
2D nuclear Overhauser enhancement spectroscopy
- NOESY-HSQC
3D NOESY relayed HSQC
- TOCSY
2D total correlation spectroscopy
- TPPI
time-proportional phase incrementation
- TSP
trimethyl silyl propionate 相似文献
16.
Jingzhaotoxin-I (JZTX-I) purified from the venom of the spider Chilobrachys jingzhao is a novel neurotoxin preferentially inhibiting cardiac sodium channel inactivation by binding to receptor site 3.The structure of this toxin in aqueous solution was investigated using 2-D ^1H-NMR techniques. The complete sequence-specific assignments of proton resonance in the ^1H-NMR spectra of JZTX-I were obtained by analyzing a series of 2-D spectra, including DQF-COSY, TOCSY and NOESY spectra, in n20 and D20. All the backbone protons except for Gin4 and more than 95% of the side-chain protons were identified by dαN,dαδ, dβN and dNN connectivities in the NOESY spectrum. These studies provide a basis for the furtherdeter mination of the solution conformation of JZTX-I. Furthermore, the secondary structure of JZTX-I was identified from NMR data. It consists mainly of a short triple-stranded antiparallel β-sheet with Trp7-Cys9, Phe20-Lys23 and Leu28-Trp31. The characteristics of the secondary structure of JZTX-I are similar to those of huwentoxin-I (HWTX-I) and hainantoxin-IV (HNTX-IV), whose structures in solution havepre viously been reported. 相似文献
17.
Glenn L. Millhauser Chris J. Stenland Kimberly A. Bolin Frank J. M. van de Ven 《Journal of biomolecular NMR》1996,7(4):331-334
Summary Alanine-rich peptides serve as models for exploring the factors that control helix structure in peptides and proteins. Scalar CH-NH couplings (3JHN) are an extremely useful measure of local helix content; however, the large alanine content in these peptides leads to significant signal overlap in the CH region of 1H 2D NMR spectra. Quantitative determination of all possible 3JHN values is, therefore, very challenging. Szyperski and co-workers [(1992) J. Magn. Reson., 99, 552–560] have recently developed a method for determining 3JHN from NOESY spectra. Because 3JHN may be determined from 2D peaks outside of the CH region, there is a much greater likelihood of identifying resolved resonances and measuring the associated coupling constants. It is demonstrated here that 3JHN can be obtained for every residue in the helical peptide Ac-(AAAAK)3A-NH2. The resulting 3JHN profile clearly identifies a helical structure in the middle of the peptide and further suggests that the respective helix termini unfold via distinct pathways.Abbreviations
3JHN
three-bond CH-NH scalar coupling constant
- NOE
nuclear Overhauser enhancement
- NOESY
two-dimensional nuclear Overhauser spectroscopy
- COSY
two-dimensional correlated spectroscopy
- DQF-COSY
two-dimensional double-quantum-filtered correlated spectroscopy
- TOCSY
two-dimensional total correlation spectroscopy
To whom correspondence should be addressed.Deceased March 5, 1996. 相似文献
18.
C. A.G. Haasnoot U. K. Pandit C. Kruk C. W. Hilbers 《Journal of biomolecular structure & dynamics》2013,31(2):449-467
Abstract 1H-NMR spectra of bleomycin A2 recorded at 500 MHz in D2O and H2O at 24°C and 3°C were investigated. Resonances of the individual spin systems were identified by using two-dimensional correlated spectroscopy (COSY), two-dimensional spin echo correlated spectroscopy (SECSY) and by the application of two-dimensional Nuclear Overhauser Effect spectroscopy (NOESY). Employment of these techniques allowed the assignment of 13 exchangeable and 59 non-exchangeable protons in the 1H NMR spectrum of bleomycin A2. By means of 2D NOE spectroscopy also interresidual connectivities could be observed. Comparison of the NOESY spectra at 3°C and 24°C suggest that at low temperatures the central part of the bleomycin A2 molecule tends to adopt an extended conformation. 相似文献
19.
Thomas Szyperski Gerhard Wider John H. Bushweller Kurt Wüthrich 《Journal of biomolecular NMR》1993,3(1):127-132
Summary The pulse sequence of a new constant-time 3D triple-resonance experiment, ct-HA[CAN]HN, is presented. This experiment delineates exclusively scalar connectivities and uses 13C–15N heteronuclear two-spin coherence to overlay the chemical shift evolution periods of the 13C and 15N nuclei, thereby providing the four resonance frequencies of the -proton, the -carbon, the amide nitrogen, and the amide proton of a given amino acid residue in three dimensions. This experiment promises to be a valid alternative to 4D experiments, providing the same information on intraresidue polypeptide backbone connectivities in 13C-15N-double-labeled proteins.Abbreviations 3D, 4D
three-dimensional, four-dimensional
- TPPI
time-proportional phase incrementation
- ct
constant-time
- rf
radiofrequency
- NOE
nuclear Overhauser enhancement
- NOESY
two-dimensional nuclear Overhauser enhancement spectroscopy
- glutaredoxin(C14S)
mutant E. coli glutaredoxin with the cysteine at position 14 replaced by serine 相似文献
20.
Summary A new program, ASNO (ASsign NOes), for computer-supported NOE cross-peak assignments is described. ASNO is used for structure refinement in several rounds of NOESY cross-peak assignments and 3D structure calculations, where the preliminary structures are used as a reference to resolve ambiguities in NOE assignments which are otherwise based on the chemical shifts available from the sequence-specific resonance assignments. The practical use of ASNO for proteins is illustrated with the structure determination of Dendrotoxin K from Dendroaspis polylepis polylepis.Abbreviations Toxin K
dendrotoxin K (or trypsin inhibitor homologue K) from the venom of the black mamba Dendroaspis polylepis polylepis
- NOE
nuclear Overhauser effect
- NOESY
NOE spectroscopy
- REDAC
use of redundant dihedral angle constraints
- RMSD
root-mean-square deviation
To whom correspondence should be addressed. 相似文献