Membrane-protein structure determination by solid-state NMR spectroscopy of microcrystals.

Abstract:

:Membrane proteins are largely underrepresented among available atomic-resolution structures. The use of detergents in protein purification procedures hinders the formation of well-ordered crystals for X-ray crystallography and leads to slower molecular tumbling, impeding the application of solution-state NMR. Solid-state magic-angle spinning NMR spectroscopy is an emerging method for membrane-protein structural biology that can overcome these technical problems. Here we present the solid-state NMR structure of the transmembrane domain of the Yersinia enterocolitica adhesin A (YadA). The sample was derived from crystallization trials that yielded only poorly diffracting microcrystals. We solved the structure using a single, uniformly (13)C- and (15)N-labeled sample. In addition, solid-state NMR allowed us to acquire information on the flexibility and mobility of parts of the structure, which, in combination with evolutionary conservation information, presents new insights into the autotransport mechanism of YadA.

journal_name

Nat Methods

journal_title

Nature methods

authors

Shahid SA,Bardiaux B,Franks WT,Krabben L,Habeck M,van Rossum BJ,Linke D

doi

10.1038/nmeth.2248

subject

Has Abstract

pub_date

2012-12-01 00:00:00

pages

1212-7

issue

12

eissn

1548-7091

issn

1548-7105

pii

nmeth.2248

journal_volume

9

pub_type

杂志文章
  • An FKBP destabilization domain modulates protein levels in Plasmodium falciparum.

    abstract::To enhance the repertoire of molecular tools for studying malaria parasite biology, we adapted a ligand-regulatable FKBP protein destabilization domain (ddFKBP) for use in P. falciparum. We destabilized the reporter yellow fluorescent protein (YFP) and the P. falciparum protease falcipain-2 in a ligand-reversible mann...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth1132

    authors: Armstrong CM,Goldberg DE

    更新日期:2007-12-01 00:00:00

  • A transgenic mouse for in vivo detection of endogenous labeled mRNA.

    abstract::Live-cell single mRNA imaging is a powerful tool but has been restricted in higher eukaryotes to artificial cell lines and reporter genes. We describe an approach that enables live-cell imaging of single endogenous labeled mRNA molecules transcribed in primary mammalian cells and tissue. We generated a knock-in mouse ...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.1551

    authors: Lionnet T,Czaplinski K,Darzacq X,Shav-Tal Y,Wells AL,Chao JA,Park HY,de Turris V,Lopez-Jones M,Singer RH

    更新日期:2011-02-01 00:00:00

  • Non-uniform refinement: adaptive regularization improves single-particle cryo-EM reconstruction.

    abstract::Cryogenic electron microscopy (cryo-EM) is widely used to study biological macromolecules that comprise regions with disorder, flexibility or partial occupancy. For example, membrane proteins are often kept in solution with detergent micelles and lipid nanodiscs that are locally disordered. Such spatial variability ne...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/s41592-020-00990-8

    authors: Punjani A,Zhang H,Fleet DJ

    更新日期:2020-12-01 00:00:00

  • Measurement of atom resolvability in cryo-EM maps with Q-scores.

    abstract::Cryogenic electron microscopy (cryo-EM) maps are now at the point where resolvability of individual atoms can be achieved. However, resolvability is not necessarily uniform throughout the map. We introduce a quantitative parameter to characterize the resolvability of individual atoms in cryo-EM maps, the map Q-score. ...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/s41592-020-0731-1

    authors: Pintilie G,Zhang K,Su Z,Li S,Schmid MF,Chiu W

    更新日期:2020-03-01 00:00:00

  • Analysis of the Human Protein Atlas Image Classification competition.

    abstract::Pinpointing subcellular protein localizations from microscopy images is easy to the trained eye, but challenging to automate. Based on the Human Protein Atlas image collection, we held a competition to identify deep learning solutions to solve this task. Challenges included training on highly imbalanced classes and pr...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/s41592-019-0658-6

    authors: Ouyang W,Winsnes CF,Hjelmare M,Cesnik AJ,Åkesson L,Xu H,Sullivan DP,Dai S,Lan J,Jinmo P,Galib SM,Henkel C,Hwang K,Poplavskiy D,Tunguz B,Wolfinger RD,Gu Y,Li C,Xie J,Buslov D,Fironov S,Kiselev A,Panchenko D,C

    更新日期:2019-12-01 00:00:00

  • Onco-proteogenomics: cancer proteomics joins forces with genomics.

    abstract::The complexities of tumor genomes are rapidly being uncovered, but how they are regulated into functional proteomes remains poorly understood. Standard proteomics workflows use databases of known proteins, but these databases do not capture the uniqueness of the cancer transcriptome, with its point mutations, unusual ...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.3138

    authors: Alfaro JA,Sinha A,Kislinger T,Boutros PC

    更新日期:2014-11-01 00:00:00

  • More options for gene editing.

    abstract::Engineering precise genetic changes in a genome is powerful way to study gene function, and several recent papers describe new applications of gene-editing tools. Working with researchers at Sangamo BioSciences, Howard Hughes Medical Institute investigator Barbara Meyer and her colleagues at the University of Californ...

    journal_title:Nature methods

    pub_type: 评论,杂志文章

    doi:10.1038/nmeth.1683

    authors: Baker M

    更新日期:2011-09-01 00:00:00

  • Rational design of true monomeric and bright photoactivatable fluorescent proteins.

    abstract::Monomeric (m)Eos2 is an engineered photoactivatable fluorescent protein widely used for super-resolution microscopy. We show that mEos2 forms oligomers at high concentrations and forms aggregates when labeling membrane proteins, limiting its application as a fusion partner. We solved the crystal structure of tetrameri...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.2021

    authors: Zhang M,Chang H,Zhang Y,Yu J,Wu L,Ji W,Chen J,Liu B,Lu J,Liu Y,Zhang J,Xu P,Xu T

    更新日期:2012-05-13 00:00:00

  • Rapid, optimized interactomic screening.

    abstract::We must reliably map the interactomes of cellular macromolecular complexes in order to fully explore and understand biological systems. However, there are no methods to accurately predict how to capture a given macromolecular complex with its physiological binding partners. Here, we present a screening method that com...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.3395

    authors: Hakhverdyan Z,Domanski M,Hough LE,Oroskar AA,Oroskar AR,Keegan S,Dilworth DJ,Molloy KR,Sherman V,Aitchison JD,Fenyö D,Chait BT,Jensen TH,Rout MP,LaCava J

    更新日期:2015-06-01 00:00:00

  • Imaging brain electric signals with genetically targeted voltage-sensitive fluorescent proteins.

    abstract::Cortical information processing relies on synaptic interactions between diverse classes of neurons with distinct electrophysiological and connection properties. Uncovering the operational principles of these elaborate circuits requires the probing of electrical activity from selected populations of defined neurons. He...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.1479

    authors: Akemann W,Mutoh H,Perron A,Rossier J,Knöpfel T

    更新日期:2010-08-01 00:00:00

  • Large-scale electrophysiology with polymer-based electrodes.

    abstract:: ...

    journal_title:Nature methods

    pub_type: 评论,杂志文章

    doi:10.1038/s41592-019-0315-0

    authors: Vogt N

    更新日期:2019-02-01 00:00:00

  • Molecular engineering: Unnatural design.

    abstract::Researchers designed an enzyme to carry out the Diels-Alder reaction, an activity not found in nature. ...

    journal_title:Nature methods

    pub_type: 评论,杂志文章

    doi:10.1038/nmeth0910-671

    authors: Doerr A

    更新日期:2010-09-01 00:00:00

  • Structural biology: 'seeing' crystals the XFEL way.

    abstract::X-ray free-electron lasers (XFELs) offer opportunities beyond classic X-ray crystallography, particularly for proteins that are difficult to crystallize. ...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.3070

    authors: Marx V

    更新日期:2014-09-01 00:00:00

  • FRETting for a more detailed interactome.

    abstract::A quantitative high-throughput FRET-based method of screening fluorescent protein fusion libraries brings the promise of more detailed interactome maps. ...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth0207-112b

    authors: Kaganman I

    更新日期:2007-02-01 00:00:00

  • An in vivo multiplexed small-molecule screening platform.

    abstract::Phenotype-based small-molecule screening is a powerful method to identify molecules that regulate cellular functions. However, such screens are generally performed in vitro under conditions that do not necessarily model complex physiological conditions or disease states. Here, we use molecular cell barcoding to enable...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.3992

    authors: Grüner BM,Schulze CJ,Yang D,Ogasawara D,Dix MM,Rogers ZN,Chuang CH,McFarland CD,Chiou SH,Brown JM,Cravatt BF,Bogyo M,Winslow MM

    更新日期:2016-10-01 00:00:00

  • A general design strategy for protein-responsive riboswitches in mammalian cells.

    abstract::RNAs are ideal for the design of gene switches that can monitor and program cellular behavior because of their high modularity and predictable structure-function relationship. We have assembled an expression platform with an embedded modular ribozyme scaffold that correlates self-cleavage activity of designer ribozyme...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.3136

    authors: Ausländer S,Stücheli P,Rehm C,Ausländer D,Hartig JS,Fussenegger M

    更新日期:2014-11-01 00:00:00

  • Visualization and analysis of single-cell RNA-seq data by kernel-based similarity learning.

    abstract::We present single-cell interpretation via multikernel learning (SIMLR), an analytic framework and software which learns a similarity measure from single-cell RNA-seq data in order to perform dimension reduction, clustering and visualization. On seven published data sets, we benchmark SIMLR against state-of-the-art met...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.4207

    authors: Wang B,Zhu J,Pierson E,Ramazzotti D,Batzoglou S

    更新日期:2017-04-01 00:00:00

  • Rapid selection of transgenic C. elegans using antibiotic resistance.

    abstract::Caenorhabditis elegans is an important model organism in biology, but until now no antibiotic selection markers have been successfully demonstrated for this species. We have developed a selection system using puromycin that allows the rapid and easy isolation of large populations of transgenic worms. This approach is ...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.1495

    authors: Semple JI,Garcia-Verdugo R,Lehner B

    更新日期:2010-09-01 00:00:00

  • Quantitative super-resolution imaging with qPAINT.

    abstract::Counting molecules in complexes is challenging, even with super-resolution microscopy. Here, we use the programmable and specific binding of dye-labeled DNA probes to count integer numbers of targets. This method, called quantitative points accumulation in nanoscale topography (qPAINT), works independently of dye phot...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.3804

    authors: Jungmann R,Avendaño MS,Dai M,Woehrstein JB,Agasti SS,Feiger Z,Rodal A,Yin P

    更新日期:2016-05-01 00:00:00

  • Gas cluster ion beam SEM for imaging of large tissue samples with 10 nm isotropic resolution.

    abstract::We demonstrate gas cluster ion beam scanning electron microscopy (SEM), in which wide-area ion milling is performed on a series of thick tissue sections. This three-dimensional electron microscopy technique acquires datasets with <10 nm isotropic resolution of each section, and these can then be stitched together to s...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/s41592-019-0641-2

    authors: Hayworth KJ,Peale D,Januszewski M,Knott GW,Lu Z,Xu CS,Hess HF

    更新日期:2020-01-01 00:00:00

  • Site-specific recombination in human embryonic stem cells induced by cell-permeant Cre recombinase.

    abstract::The biomedical application of human embryonic stem (hES) cells will increasingly depend on the availability of technologies for highly controlled genetic modification. In mouse genetics, conditional mutagenesis using site-specific recombinases has become an invaluable tool for gain- and loss-of-function studies. Here ...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth884

    authors: Nolden L,Edenhofer F,Haupt S,Koch P,Wunderlich FT,Siemen H,Brüstle O

    更新日期:2006-06-01 00:00:00

  • Tracking transmitter-gated P2X cation channel activation in vitro and in vivo.

    abstract::We present a noninvasive approach to track activation of ATP-gated P2X receptors and potentially other transmitter-gated cation channels that show calcium fluxes. We genetically engineered rat P2X receptors to carry calcium sensors near the channel pore and tested this as a reporter for P2X(2) receptor opening. The me...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth1144

    authors: Richler E,Chaumont S,Shigetomi E,Sagasti A,Khakh BS

    更新日期:2008-01-01 00:00:00

  • The inside tag.

    abstract::An uncharged CoA precursor that can enter the cell is used for covalent, site-specific labeling of proteins inside living cells. ...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth1106-872b

    authors: Kaganman I

    更新日期:2006-11-01 00:00:00

  • Putative cell type discovery from single-cell gene expression data.

    abstract::We present the Single-Cell Clustering Assessment Framework, a method for the automated identification of putative cell types from single-cell RNA sequencing (scRNA-seq) data. By iteratively applying a machine learning approach to a given set of cells, we simultaneously identify distinct cell groups and a weighted list...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/s41592-020-0825-9

    authors: Miao Z,Moreno P,Huang N,Papatheodorou I,Brazma A,Teichmann SA

    更新日期:2020-06-01 00:00:00

  • Genetic incorporation of unnatural amino acids into proteins in mammalian cells.

    abstract::We developed a general approach that allows unnatural amino acids with diverse physicochemical and biological properties to be genetically encoded in mammalian cells. A mutant Escherichia coli aminoacyl-tRNA synthetase (aaRS) is first evolved in yeast to selectively aminoacylate its tRNA with the unnatural amino acid ...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth1016

    authors: Liu W,Brock A,Chen S,Chen S,Schultz PG

    更新日期:2007-03-01 00:00:00

  • Improved and expanded Q-system reagents for genetic manipulations.

    abstract::The Q system is a repressible binary expression system for transgenic manipulations in living organisms. Through protein engineering and in vivo functional tests, we report here variants of the Q-system transcriptional activator, including QF2, for driving strong and ubiquitous expression in all Drosophila tissues. Ou...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.3250

    authors: Riabinina O,Luginbuhl D,Marr E,Liu S,Wu MN,Luo L,Potter CJ

    更新日期:2015-03-01 00:00:00

  • Whole-genome genotyping with the single-base extension assay.

    abstract::We describe an efficient, accurate and robust whole-genome genotyping (WGG) assay based on a two-color, single-base extension (SBE), single-nucleotide polymorphism (SNP)-scoring step. We report genotyping results for biallelic International HapMap quality control (QC) SNPs using a single probe per locus. We show scala...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth842

    authors: Steemers FJ,Chang W,Lee G,Barker DL,Shen R,Gunderson KL

    更新日期:2006-01-01 00:00:00

  • Visual proteomics of the human pathogen Leptospira interrogans.

    abstract::Systems biology conceptualizes biological systems as dynamic networks of interacting elements, whereby functionally important properties are thought to emerge from the structure of such networks. Owing to the ubiquitous role of complexes of interacting proteins in biological systems, their subunit composition and temp...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.1390

    authors: Beck M,Malmström JA,Lange V,Schmidt A,Deutsch EW,Aebersold R

    更新日期:2009-11-01 00:00:00

  • Two-color, two-photon uncaging of glutamate and GABA.

    abstract::We developed a caged GABA (gamma-aminobutyric acid), which, when combined with an appropriate caged glutamate, allows bimodal control of neuronal membrane potential with subcellular resolution using optically independent two-photon uncaging of each neurotransmitter. We used two-color, two-photon uncaging to fire and b...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.1413

    authors: Kantevari S,Matsuzaki M,Kanemoto Y,Kasai H,Ellis-Davies GC

    更新日期:2010-02-01 00:00:00

  • Finding S-sulfinylated proteins.

    abstract:: ...

    journal_title:Nature methods

    pub_type: 评论,杂志文章

    doi:10.1038/s41592-018-0200-2

    authors: Doerr A

    更新日期:2018-11-01 00:00:00