当前位置： SCI文献检索 > JOURNAL OF BIOMOLECULAR NMR期刊下所有文献 > Improved segmental isotope labeling of proteins and application to a larger protein.

Improved segmental isotope labeling of proteins and application to a larger protein.

Abstract：

:A new isotope labeling technique for peptide segments in a protein sample was recently established using the protein splicing element intein [Yamazaki et al. (1998) J. Am. Chem. Soc., 120, 5591-5592]. This method makes it possible to observe signals of a selected amino (N-) or carboxyl (C-) terminal region along a peptide chain. However, there is a problem with the yield of the segmentally labeled protein. In this paper, we report an increase in the yield of the protein that enables the production of sufficient amounts of segmentally 13C/15N-labeled protein samples. This was achieved by improvement of the expression level of the N-terminal fragment in cells and the efficiency of refolding into the active splicing conformation. The N-terminal fragment was expressed as a fused protein with the cellulose binding domain at its N-terminus, which was expressed as an insoluble peptide in cells and the expression level was increased. Incubation with 2.5 M urea and 50% glycerol increased the efficiency of the refolding greatly, thereby raising the final yields of the ligated proteins. The feasibility of application of the method to a high-molecular-weight protein was demonstrated by the results for a maltose binding protein consisting of 370 amino acids. All four examined joints in the maltose binding protein were successfully ligated to produce segmentally labeled protein samples.

journal_name

J Biomol NMR

journal_title

Journal of biomolecular NMR

authors

Otomo T,Teruya K,Uegaki K,Yamazaki T,Kyogoku Y

doi

10.1023/a:1008308128050

subject

Has Abstract

pub_date

1999-06-01 00:00:00

pages

105-14

issue

2

eissn

0925-2738

issn

1573-5001

journal_volume

14

pub_type

杂志文章

相关文献

JOURNAL OF BIOMOLECULAR NMR文献大全
  • NMR of bicelles: orientation and mosaic spread of the liquid-crystal director under sample rotation.

    abstract：:Model-membrane systems composed of liquid-crystalline bicellar phases can be uniaxially oriented with respect to a magnetic field, thereby facilitating structural and dynamics studies of membrane-associated proteins. Here we quantitatively characterize a method that allows the manipulation of the direction of this uni...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1023/a:1022236217018

    authors： Zandomeneghi G,Tomaselli M,Williamson PT,Meier BH

    更新日期：2003-02-01 00:00:00

  • Validation of a new restraint docking method for solution structure determinations of protein-ligand complexes.

    abstract：:A new method is proposed for docking ligands into proteins in cases where an NMR-determined solution structure of a related complex is available. The method uses a set of experimentally determined values for protein-ligand, ligand-ligand, and protein-protein restraints for residues in or near to the binding site, comb...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1023/a:1008379225053

    authors： Polshakov VI,Morgan WD,Birdsall B,Feeney J

    更新日期：1999-06-01 00:00:00

  • Quantifying Lipari-Szabo modelfree parameters from 13CO NMR relaxation experiments.

    abstract：:It is proposed to obtain effective Lipari-Szabo order parameters and local correlation times for relaxation vectors of protein (13)CO nuclei by carrying out a (13)CO-R(1) auto relaxation experiment, a transverse (13)CO CSA/13CO-13Calpha CSA/dipolar cross correlation and a transverse (13)CO CSA/(13)CO-(15)N CSA/dipolar...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-006-9047-4

    authors： Wang T,Weaver DS,Cai S,Zuiderweg ER

    更新日期：2006-10-01 00:00:00

  • Structure and conformation in solution of the parallel-stranded hybrid alpha-d(CGCAATTCGC).beta-d(GCGTTAAGCG) by high-resolution 2D NMR.

    abstract：:The unnatural duplex oligonucleotide alpha-d(CGCAATTCGC).beta-d(GCGTTAAGCG) was analyzed by high-resolution NMR methods. All of the exchangeable imino and nonexchangeable protons of the duplex were assigned. Detection of all 10 of the exchangeable imino protons confirms that a parallel, unsymmetrical duplex is formed....

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/BF01875321

    authors： Gmeiner WH,Rayner B,Morvan F,Imbach JL,Lown JW

    更新日期：1992-05-01 00:00:00

  • Determination of the solution structure of the SH3 domain of human p56 Lck tyrosine kinase.

    abstract：:The solution structure of the SH3 domain of human p56 Lck tyrosine kinase (Lck-SH3) has been determined by multidimensional heteronuclear NMR spectroscopy. The structure was calculated from a total of 935 experimental restraints comprising 785 distance restraints derived from 1017 assigned NOE cross peaks and 150 dihe...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/BF00211158

    authors： Hiroaki H,Klaus W,Senn H

    更新日期：1996-09-01 00:00:00

  • Automated 1H and 13C chemical shift prediction using the BioMagResBank.

    abstract：:A computer program has been developed to accurately and automatically predict the 1H and 13C chemical shifts of unassigned proteins on the basis of sequence homology. The program (called SHIFTY) uses standard sequence alignment techniques to compare the sequence of an unassigned protein against the BioMagResBank--a pu...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1023/a:1018373822088

    authors： Wishart DS,Watson MS,Boyko RF,Sykes BD

    更新日期：1997-12-01 00:00:00

  • Comparison of the backbone dynamics of wild-type Hydrogenobacter thermophilus cytochrome c(552) and its b-type variant.

    abstract：:Cytochrome c552 from the thermophilic bacterium Hydrogenobacter thermophilus is a typical c-type cytochrome which binds heme covalently via two thioether bonds between the two heme vinyl groups and two cysteine thiol groups in a CXXCH sequence motif. This protein was converted to a b-type cytochrome by substitution of...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-015-9938-3

    authors： Tozawa K,Ferguson SJ,Redfield C,Smith LJ

    更新日期：2015-06-01 00:00:00

  • Spectral editing: selection of methyl groups in multidimensional solid-state magic-angle spinning NMR.

    abstract：:A simple spectroscopic filtering technique is presented that may aid the assignment of (13)C and (15)N resonances of methyl-containing amino-acids in solid-state magic-angle spinning (MAS) NMR. A filtering block that selects methyl resonances is introduced in two-dimensional (2D) (13)C-homonuclear and (15)N-(13)C hete...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-006-9078-x

    authors： Jehle S,Hiller M,Rehbein K,Diehl A,Oschkinat H,van Rossum BJ

    更新日期：2006-11-01 00:00:00

  • Probing non-specific interactions of Ca²⁺-calmodulin in E. coli lysate.

    abstract：:The biological environment in which a protein performs its function is a crowded milieu containing millions of molecules that can potentially lead to a great many transient, non-specific interactions. NMR spectroscopy is especially well suited to study these weak molecular contacts. Here, non-specific interactions bet...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-013-9705-2

    authors： Latham MP,Kay LE

    更新日期：2013-03-01 00:00:00

  • Solid-state NMR, electrophysiology and molecular dynamics characterization of human VDAC2.

    abstract：:The voltage-dependent anion channel (VDAC) is the most abundant protein of the outer mitochondrial membrane and constitutes the major pathway for the transport of ADP, ATP, and other metabolites. In this multidisciplinary study we combined solid-state NMR, electrophysiology, and molecular dynamics simulations, to stud...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-014-9876-5

    authors： Gattin Z,Schneider R,Laukat Y,Giller K,Maier E,Zweckstetter M,Griesinger C,Benz R,Becker S,Lange A

    更新日期：2015-04-01 00:00:00

  • GFT projection NMR based resonance assignment of membrane proteins: application to subunit C of E. coli F(1)F (0) ATP synthase in LPPG micelles.

    abstract：:G-matrix FT projection NMR spectroscopy was employed for resonance assignment of the 79-residue subunit c of the Escherichia coli F(1)F(0) ATP synthase embedded in micelles formed by lyso palmitoyl phosphatidyl glycerol (LPPG). Five GFT NMR experiments, that is, (3,2)D HNNCO, L-(4,3)D HNNC (alphabeta) C (alpha), L-(4,...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-008-9224-8

    authors： Zhang Q,Atreya HS,Kamen DE,Girvin ME,Szyperski T

    更新日期：2008-03-01 00:00:00

  • Sequential correlation of anomeric ribose protons and intervening phosphorus in RNA oligonucleotides by a 1H, 13C, 31P triple resonance experiment: HCP-CCH-TOCSY.

    abstract：:A three-dimensional 1H, 13C, 31P triple resonance experiment, HCP-CCH-TOCSY, is presented which provides unambiguous through-bond correlation of all 1H ribose protons on the 5' and 3' sides of the intervening phosphorus along the backbone bonding network in 13C-labeled RNA oligonucleotides. The correlation of the comp...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/BF00227473

    authors： Marino JP,Schwalbe H,Anklin C,Bermel W,Crothers DM,Griesinger C

    更新日期：1995-01-01 00:00:00

  • Two-dimensional NMR lineshape analysis of single, multiple, zero and double quantum correlation experiments.

    abstract：:NMR spectroscopy provides a powerful approach for the characterisation of chemical exchange and molecular interactions by analysis of series of experiments acquired over the course of a titration measurement. The appearance of NMR resonances undergoing chemical exchange depends on the frequency difference relative to ...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-019-00297-7

    authors： Waudby CA,Ouvry M,Davis B,Christodoulou J

    更新日期：2020-01-01 00:00:00

  • Triple resonance ¹⁵Ν NMR relaxation experiments for studies of intrinsically disordered proteins.

    abstract：:Description of protein dynamics is known to be essential in understanding their function. Studies based on a well established [Formula: see text] NMR relaxation methodology have been applied to a large number of systems. However, the low dispersion of [Formula: see text] chemical shifts very often observed within intr...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-017-0138-1

    authors： Srb P,Nováček J,Kadeřávek P,Rabatinová A,Krásný L,Žídková J,Bobálová J,Sklenář V,Žídek L

    更新日期：2017-11-01 00:00:00

  • PROSHIFT: protein chemical shift prediction using artificial neural networks.

    abstract：:The importance of protein chemical shift values for the determination of three-dimensional protein structure has increased in recent years because of the large databases of protein structures with assigned chemical shift data. These databases have allowed the investigation of the quantitative relationship between chem...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1023/a:1023060720156

    authors： Meiler J

    更新日期：2003-05-01 00:00:00

  • Determination of protein global folds using backbone residual dipolar coupling and long-range NOE restraints.

    abstract：:We report the determination of the global fold of human ubiquitin using protein backbone NMR residual dipolar coupling and long-range nuclear Overhauser effect (NOE) data as conformational restraints. Specifically, by use of a maximum of three backbone residual dipolar couplings per residue (Ni-H N i, Ni-C'(i-1), H N ...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1023/a:1021954812977

    authors： Giesen AW,Homans SW,Brown JM

    更新日期：2003-01-01 00:00:00

  • The secondary structure of a pyrimidine-guanine sequence-specific ribonuclease possessing cytotoxic activity from the oocytes of Rana catesbeiana.

    abstract：: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 ...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/BF00410331

    authors： Chen C,Hom K,Huang RF,Chou PJ,Liao YD,Huang T

    更新日期：1996-10-01 00:00:00

  • Site-specific labeling of proteins with NMR-active unnatural amino acids.

    abstract：:A large number of amino acids other than the canonical amino acids can now be easily incorporated in vivo into proteins at genetically encoded positions. The technology requires an orthogonal tRNA/aminoacyl-tRNA synthetase pair specific for the unnatural amino acid that is added to the media while a TAG amber or frame...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章,评审

    doi：10.1007/s10858-009-9365-4

    authors： Jones DH,Cellitti SE,Hao X,Zhang Q,Jahnz M,Summerer D,Schultz PG,Uno T,Geierstanger BH

    更新日期：2010-01-01 00:00:00

  • (1)H, (13)C and (15)N NMR backbone assignments of the 269-residue serine protease PB92 from Bacillus alcalophilus.

    abstract：:The (1)H, (13)C and (15)N NMR resonances of the backbone of serine protease PB92 have been assigned. This 269-residue protein is one of the largest monomeric proteins assigned so far. The amount and quality of information available suggest that even larger proteins could be assigned with present methods. Measured chem...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/BF00178340

    authors： Fogh RH,Schipper D,Boelens R,Kaptein R

    更新日期：1994-01-01 00:00:00

  • Investigation of backbone dynamics and local geometry of bio-molecules using calculated NMR chemical shifts and anisotropies.

    abstract：:Prerequisite for chemical shift (CS) and CS tensor calculations are highly refined structures defining the molecular surroundings of the nuclei under study. Here, we present geometry optimizations with 13C and 15N CS constraints for large bio-molecules like peptides and proteins. The method discussed here provides bot...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-019-00284-y

    authors： Sternberg U,Witter R

    更新日期：2019-12-01 00:00:00

  • 1H-1H correlations across N-H...N hydrogen bonds in nucleic acids.

    abstract：:In 2HJ(NN)-COSY experiments, which correlate protons with donor/acceptor nitrogens across Nd...HNa bonds, the receptor nitrogen needs to be assigned in order to unambiguously identify the hydrogen bond. For many situations this is a non-trivial task which is further complicated by poor dispersion of (Na,Nd) resonances...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1023/a:1013340227140

    authors： Majumdar A,Gosser Y,Patel DJ

    更新日期：2001-12-01 00:00:00

  • Practical considerations over spectral quality in solid state NMR spectroscopy of soluble proteins.

    abstract：:Great theoretical and methodological advances are pushing the limits of resolution and sensitivity in solid state NMR (SSNMR). However, sample preparation remains a critical issue for the success of an experiment. The factors affecting spectral quality in SSNMR samples are discussed, examining cases encountered in the...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-013-9776-0

    authors： Fragai M,Luchinat C,Parigi G,Ravera E

    更新日期：2013-10-01 00:00:00

  • Extension of the HA-detection based approach: (HCA)CON(CA)H and (HCA)NCO(CA)H experiments for the main-chain assignment of intrinsically disordered proteins.

    abstract：:Extensive resonance overlap exacerbates assignment of intrinsically disordered proteins (IDPs). This issue can be circumvented by utilizing (15)N, (13)C' and (1)H(N) spins, where the chemical shift dispersion is mainly dictated by the characteristics of consecutive amino acid residues. Especially (15)N and (13)C' spin...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-011-9470-z

    authors： Mäntylahti S,Hellman M,Permi P

    更新日期：2011-02-01 00:00:00

  • Maximum entropy approach to the determination of solution conformation of flexible polypeptides by global conformational analysis and NMR spectroscopy--application to DNS1-c-[D-A2,bu2,Trp4,Leu5]enkephalin and DNS1-c-[D-A2bu2,Trp4,D-Leu5]enkephalin.

    abstract：:A method is proposed to determine the conformational equilibrium of flexible polypeptides in solution, using the data provided by NMR spectroscopy and theoretical conformational calculations. The algorithm consists of the following three steps: (i) search of the conformational space in order to find conformations with...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1023/a:1008349424452

    authors： Groth M,Malicka J,Czaplewski C,Ołdziej S,Lankiewicz L,Wiczk W,Liwo A

    更新日期：1999-12-01 00:00:00

  • 1H(C) and 1H(N) total NOE correlations in a single 3D NMR experiment. 15N and 13C time-sharing in t1 and t2 dimensions for simultaneous data acquisition.

    abstract：:Simultaneous data acquisition in time-sharing (TS) multi-dimensional NMR experiments has been shown an effective means to reduce experimental time, and thus to accelerate structure determination of proteins. This has been accomplished by spin evolution time-sharing of the X and Y heteronuclei, such as (15)N and (13)C,...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1023/a:1025407905478

    authors： Xia Y,Yee A,Arrowsmith CH,Gao X

    更新日期：2003-11-01 00:00:00

  • Accessible surface area from NMR chemical shifts.

    abstract：:Accessible surface area (ASA) is the surface area of an atom, amino acid or biomolecule that is exposed to solvent. The calculation of a molecule's ASA requires three-dimensional coordinate data and the use of a "rolling ball" algorithm to both define and calculate the ASA. For polymers such as proteins, the ASA for i...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-015-9957-0

    authors： Hafsa NE,Arndt D,Wishart DS

    更新日期：2015-07-01 00:00:00

  • Attachment of an NMR-invisible solubility enhancement tag using a sortase-mediated protein ligation method.

    abstract：:Sample solubility is essential for structural studies of proteins by solution NMR. Attachment of a solubility enhancement tag, such as GB1, MBP and thioredoxin, to a target protein has been used for this purpose. However, signal overlap of the tag with the target protein often made the spectral analysis difficult. Her...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-008-9296-5

    authors： Kobashigawa Y,Kumeta H,Ogura K,Inagaki F

    更新日期：2009-03-01 00:00:00

  • NMRlib: user-friendly pulse sequence tools for Bruker NMR spectrometers.

    abstract：:We present NMRlib, a suite of jython-based tools designed for Bruker spectrometers (TopSpin versions 3.2-4.0) that allow easy setup, management, and exchange of NMR experiments. A NMR experiment can be set up and executed in a few clicks by navigating through the NMRlib GUI tree structure, without any further paramete...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-019-00249-1

    authors： Favier A,Brutscher B

    更新日期：2019-05-01 00:00:00

  • Solution structure of the N-terminal domain of the Staphylococcus aureus hibernation promoting factor.

    abstract：:Staphylococcus aureus hibernation promoting factor (SaHPF) is a 22,2 kDa protein which plays a crucial role in 100S Staphylococcus aureus ribosome formation during stress. SaHPF consists of N-terminal domain (NTD) that prevents proteins synthesis by binding to the 30S subunit at the P- and A-sites, connected through a...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-019-00254-4

    authors： Usachev KS,Validov SZ,Khusainov IS,Varfolomeev AA,Klochkov VV,Aganov AV,Yusupov MM

    更新日期：2019-05-01 00:00:00

  • Nitrogen detected TROSY at high field yields high resolution and sensitivity for protein NMR.

    abstract：:Detection of (15)N in multidimensional NMR experiments of proteins has sparsely been utilized because of the low gyromagnetic ratio (γ) of nitrogen and the presumed low sensitivity of such experiments. Here we show that selecting the TROSY components of proton-attached (15)N nuclei (TROSY (15)NH) yields high quality s...

    journal_title：Journal of biomolecular NMR

    pub_type： 杂志文章

    doi：10.1007/s10858-015-9991-y

    authors： Takeuchi K,Arthanari H,Shimada I,Wagner G

    更新日期：2015-12-01 00:00:00