Abstract：
:Gap junction channels assembled from connexin protein subunits mediate intercellular transfer of ions and metabolites. Impaired channel function is implicated in several hereditary human diseases. In particular, defective permeation of cAMP or inositol-1,4,5-trisphosphate (InsP(3)) through connexin channels is associated with peripheral neuropathies and deafness, respectively. Here we present a method to estimate the permeability of single gap junction channels to second messengers. Using HeLa cells that overexpressed wild-type human connexin 26 (HCx26wt) as a model system, we combined measurements of junctional conductance and fluorescence resonance energy transfer (FRET) emission ratio of biosensors selective for cAMP and InsP(3). The unitary permeabilities to cAMP (47 x 10(-3) +/- 15 x 10(-3) microm(3)/s) and InsP(3) (60 x 10(-3) +/- 12 x 10(-3) microm(3)/s) were similar, but substantially larger than the unitary permeability to lucifer yellow (LY; 7 +/- 3 x 10(-3) microm(3)/s), an exogenous tracer. This method permits quantification of defects of metabolic coupling and can be used to investigate interdependence of intercellular diffusion and cross-talk between diverse signaling pathways.
journal_name
Nat Methodsjournal_title
Nature methodsauthors
Hernandez VH,Bortolozzi M,Pertegato V,Beltramello M,Giarin M,Zaccolo M,Pantano S,Mammano Fdoi
10.1038/nmeth1031subject
Has Abstractpub_date
2007-04-01 00:00:00pages
353-8issue
4eissn
1548-7091issn
1548-7105pii
nmeth1031journal_volume
4pub_type
杂志文章相关文献
NATURE METHODS文献大全abstract：:Shotgun sequencing enables the reconstruction of genomes from complex microbial communities, but because assembly does not reconstruct entire genomes, it is necessary to bin genome fragments. Here we present CONCOCT, a new algorithm that combines sequence composition and coverage across multiple samples, to automatica...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.3103
更新日期：2014-11-01 00:00:00
abstract：:We introduce a method for sequencing peptides by mass spectrometry using a metalloendopeptidase that cleaves proteins at the amino side of lysine (Lys-N). When analyzed by electron transfer dissociation (ETD)-based mass spectrometric sequencing, Lys-N-digested peptides that contain a single lysine residue produce spec...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.1204
更新日期：2008-05-01 00:00:00
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
更新日期：2016-10-01 00:00:00
abstract：:MicroRNAs (miRNAs) play an essential role in the post-transcriptional regulation of animal development and physiology. However, in vivo studies aimed at linking miRNA function to the biology of distinct cell types within complex tissues remain challenging, partly because in vivo miRNA-profiling methods lack cellular r...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.4610
更新日期：2018-04-01 00:00:00
abstract：:Fluorescent proteins have become extremely popular tools for in vivo imaging and especially for the study of localization, motility and interaction of proteins in living cells. Here we report TagRFP, a monomeric red fluorescent protein, which is characterized by high brightness, complete chromophore maturation, prolon...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth1062
更新日期：2007-07-01 00:00:00
abstract：:We introduce field synthesis, a theorem and method that can be used to synthesize any scanned or dithered light sheet, including those used in lattice light-sheet microscopy (LLSM), from an incoherent superposition of one-dimensional intensity distributions. Compared to LLSM, this user-friendly and modular approach of...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/s41592-019-0327-9
更新日期：2019-03-01 00:00:00
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
更新日期：2014-11-01 00:00:00
abstract：:Tumors from individuals with cancer are frequently genetically profiled for information about the driving forces behind the disease. We present the CancerMine resource, a text-mined and routinely updated database of drivers, oncogenes and tumor suppressors in different types of cancer. All data are available online ( ...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/s41592-019-0422-y
更新日期：2019-06-01 00:00:00
abstract：:Recent efforts in neuroscience research have been aimed at obtaining detailed anatomical neuronal wiring maps as well as information on how neurons in these networks engage in dynamic activities. Although the entire connectivity map of the nervous system of Caenorhabditis elegans has been known for more than 25 years,...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.2637
更新日期：2013-10-01 00:00:00
abstract：:Current extracellular multisite recordings suffer from low signal-to-noise ratio, limiting the monitoring to action potentials, and preclude detection of subthreshold synaptic potentials. Here we report an approach to induce Aplysia californica neurons to actively engulf protruding microelectrodes, providing 'in-cell ...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.1420
更新日期：2010-03-01 00:00:00
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
更新日期：2010-09-01 00:00:00
abstract：:We present an easy-to-use integrated software suite, DIA-NN, that exploits deep neural networks and new quantification and signal correction strategies for the processing of data-independent acquisition (DIA) proteomics experiments. DIA-NN improves the identification and quantification performance in conventional DIA ...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/s41592-019-0638-x
更新日期：2020-01-01 00:00:00
abstract：: ...
journal_title：Nature methods
pub_type： 社论
doi：10.1038/s41592-018-0257-y
更新日期：2018-12-01 00:00:00
abstract：:We developed a method that allows release of intact membrane protein complexes from amphipols, bicelles and nanodiscs in the gas phase for observation by mass spectrometry (MS). Current methods involve release of membrane protein complexes from detergent micelles, which reveals subunit composition and lipid binding. W...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.2691
更新日期：2013-12-01 00:00:00
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
更新日期：2010-02-01 00:00:00
abstract：:Precisely defining the roles of specific cell types is an intriguing frontier in the study of intact biological systems and has stimulated the rapid development of genetically encoded tools for observation and control. However, targeting these tools with adequate specificity remains challenging: most cell types are be...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.2996
更新日期：2014-07-01 00:00:00
abstract：:Methods for genomic analysis at single-cell resolution enable new understanding of complex biological phenomena. Single-cell techniques, ranging from flow cytometry and microfluidics to PCR and sequencing, are used to understand the cellular composition of complex tissues, find new microbial species and perform genome...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth0411-311
更新日期：2011-04-01 00:00:00
abstract：:Photoconvertible fluorescent proteins are potential tools for investigating dynamic processes in living cells and for emerging super-resolution microscopy techniques. Unfortunately, most probes in this class are hampered by oligomerization, small photon budgets or poor photostability. Here we report an EosFP variant t...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.1296
更新日期：2009-02-01 00:00:00
abstract：:Focused-ion-beam scanning electron microscopy (FIB-SEM) has become an essential tool for studying neural tissue at resolutions below 10 nm × 10 nm × 10 nm, producing data sets optimized for automatic connectome tracing. We present a technical advance, ultrathick sectioning, which reliably subdivides embedded tissue sa...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.3292
更新日期：2015-04-01 00:00:00
abstract：:Transcription activator-like effector (TALE) proteins have gained broad appeal as a platform for targeted DNA recognition, largely owing to their simple rules for design. These rules relate the base specified by a single TALE repeat to the identity of two key residues (the repeat variable diresidue, or RVD) and enable...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.3330
更新日期：2015-05-01 00:00:00
abstract：:Recording light-microscopy images of large, nontransparent specimens, such as developing multicellular organisms, is complicated by decreased contrast resulting from light scattering. Early zebrafish development can be captured by standard light-sheet microscopy, but new imaging strategies are required to obtain high-...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.1476
更新日期：2010-08-01 00:00:00
abstract：:We present eXpress, a software package for efficient probabilistic assignment of ambiguously mapping sequenced fragments. eXpress uses a streaming algorithm with linear run time and constant memory use. It can determine abundances of sequenced molecules in real time and can be applied to ChIP-seq, metagenomics and oth...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.2251
更新日期：2013-01-01 00:00:00
abstract：:The commonly used, monomeric EYFP enabled imaging of intracellular protein structures beyond the optical resolution limit ('super-resolution' imaging) in living cells. By combining photoinduced activation of single EYFP fusions and time-lapse imaging, we obtained sub-40 nm resolution images of the filamentous superstr...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.1258
更新日期：2008-11-01 00:00:00
abstract：:We describe factored spectrally transformed linear mixed models (FaST-LMM), an algorithm for genome-wide association studies (GWAS) that scales linearly with cohort size in both run time and memory use. On Wellcome Trust data for 15,000 individuals, FaST-LMM ran an order of magnitude faster than current efficient algo...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.1681
更新日期：2011-09-04 00:00:00
abstract：:In mass-spectrometry-based proteomics, the identification and quantification of peptides and proteins heavily rely on sequence database searching or spectral library matching. The lack of accurate predictive models for fragment ion intensities impairs the realization of the full potential of these approaches. Here, we...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/s41592-019-0426-7
更新日期：2019-06-01 00:00:00
abstract：:The mechanical properties of the cellular microenvironment and their spatiotemporal variations are thought to play a central role in sculpting embryonic tissues, maintaining organ architecture and controlling cell behavior, including cell differentiation. However, no direct in vivo and in situ measurement of mechanica...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.4101
更新日期：2017-02-01 00:00:00
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
更新日期：2010-09-01 00:00:00
abstract：:An amendment to this paper has been published and can be accessed via a link at the top of the paper. ...
journal_title：Nature methods
pub_type： 已发布勘误
doi：10.1038/s41592-020-0733-z
更新日期：2020-02-01 00:00:00
abstract：:The normalization of RNA-seq data is essential for accurate downstream inference, but the assumptions upon which most normalization methods are based are not applicable in the single-cell setting. Consequently, applying existing normalization methods to single-cell RNA-seq data introduces artifacts that bias downstrea...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.4263
更新日期：2017-06-01 00:00:00
abstract：:Small molecules are known to stabilize membrane proteins and to modulate their function and oligomeric state, but such interactions are often hard to precisely define. Here we develop and apply a high-resolution, Orbitrap mass spectrometry-based method for analyzing intact membrane protein-ligand complexes. Using this...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.3771
更新日期：2016-04-01 00:00:00
