Abstract：
:Correct reconstruction of macromolecular structure by cryo-electron microscopy (cryo-EM) relies on accurate determination of the orientation of single-particle images. For small (<100 kDa) DNA-binding proteins, obtaining particle images with sufficiently asymmetric features to correctly guide alignment is challenging. We apply DNA origami to construct molecular goniometers-instruments that precisely orient objects-and use them to dock a DNA-binding protein on a double-helix stage that has user-programmable tilt and rotation angles. We construct goniometers with 14 different stage configurations to orient and visualize the protein just above the cryo-EM grid surface. Each goniometer has a distinct barcode pattern that we use during particle classification to assign angle priors to the bound protein. We use goniometers to obtain a 6.5-Å structure of BurrH, an 82-kDa DNA-binding protein whose helical pseudosymmetry prevents accurate image orientation using traditional cryo-EM. Our approach should be adaptable to other DNA-binding proteins as well as small proteins fused to DNA-binding domains.
journal_name
Nat Biotechnoljournal_title
Nature biotechnologyauthors
Aksel T,Yu Z,Cheng Y,Douglas SMdoi
10.1038/s41587-020-0716-8subject
Has Abstractpub_date
2020-10-19 00:00:00eissn
1087-0156issn
1546-1696pii
10.1038/s41587-020-0716-8pub_type
杂志文章abstract：:Thermostable enzymes combine catalytic specificity with the toughness required to withstand industrial reaction conditions. Stabilized enzymes also provide robust starting points for evolutionary improvement of other protein properties. We recently created a library of at least 2,300 new active chimeras of the biotech...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/nbt1333
更新日期：2007-09-01 00:00:00
abstract：:Cells isolated from the embryonic, neonatal, and adult rodent central nervous system divide in response to epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF-2), while retaining the ability to differentiate into neurons and glia. These cultures can be grown in aggregates termed neurospheres, which conta...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/88158
更新日期：2001-05-01 00:00:00
abstract：:Transplantation of pancreatic progenitors or insulin-secreting cells derived from human embryonic stem cells (hESCs) has been proposed as a therapy for diabetes. We describe a seven-stage protocol that efficiently converts hESCs into insulin-producing cells. Stage (S) 7 cells expressed key markers of mature pancreatic...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/nbt.3033
更新日期：2014-11-01 00:00:00
abstract：:A central hurdle in developing small interfering RNAs (siRNAs) as therapeutics is the inefficiency of their delivery across the plasma and endosomal membranes to the cytosol, where they interact with the RNA interference machinery. With the aim of improving endosomal release, a poorly understood and inefficient proces...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/nbt.3298
更新日期：2015-08-01 00:00:00
abstract：:Changes in glycosylation are often associated with disease progression, but the genetic and metabolic basis of these events is rarely understood in detail at a molecular level. We describe a metabolism-based approach to the selection of mutants in glycoconjugate biosynthesis that provides insight into regulatory mecha...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/89305
更新日期：2001-06-01 00:00:00
abstract：:In the version of this article initially published, a GlaxoSmithKline compound referred to as GSK298559 was listed in Table 1 as a RIPK1 inhibitor in phase 1 trials for inflammatory bowel disease. The correct designation is GSK2983559; however, this compound is a RIPK2 inhibitor and thus should not have been included ...
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pub_type： 杂志文章,已发布勘误
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pub_type： 杂志文章,评审
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pub_type： 杂志文章,评审
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journal_title：Nature biotechnology
pub_type： 杂志文章,评审
doi：10.1038/nbt.4022
更新日期：2017-12-01 00:00:00
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journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/nbt.2355
更新日期：2012-10-01 00:00:00
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pub_type： 杂志文章,评审
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更新日期：2017-03-01 00:00:00
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journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/9865
更新日期：1999-06-01 00:00:00
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journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/83489
更新日期：2001-01-01 00:00:00
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journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/13684
更新日期：1999-10-01 00:00:00
abstract：:Although mass spectrometry has become a powerful tool for the functional analysis of biological systems, complete proteome characterization cannot yet be achieved. Instead, the sheer complexity of living organisms demands fractionation of cellular extracts to enable more targeted analyses. Here, we introduce the conce...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/nbt1076
更新日期：2005-04-01 00:00:00
abstract：:Isolating antigen-specific T lymphocytes is hampered by the low frequency of the cells and the low affinity between T-cell receptors (TCR) and antigen. We describe the isolation and purification of antigen-specific CD8+ T lymphocytes from mixed T-cell populations. Magnetic beads coated with major histocompatibility co...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/nbt0398-281
更新日期：1998-03-01 00:00:00
abstract：:The oncofetal fibronectin (B-FN) isoform is present in vessels of neoplastic tissues during angiogenesis but not in mature vessels. B-FN could therefore provide a target for diagnostic imaging and therapy of cancer. Phage display libraries have been used to isolate human antibody fragments with pan-species recognition...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/nbt1197-1271
更新日期：1997-11-01 00:00:00
abstract：:The unique bread-making characteristic of wheat flour is closely related to the elasticity and extensibility of the gluten proteins stored in the starchy endosperm, particularly the high-molecular-weight glutenin subunits (HMW-GS), which are important in determining gluten and dough elasticity. The quality of wheat cu...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/nbt0996-1155
更新日期：1996-09-01 00:00:00
abstract：:Wild relatives of crops are an important source of genetic diversity for agriculture, but their gene repertoire remains largely unexplored. We report the establishment and analysis of a pan-genome of Glycine soja, the wild relative of cultivated soybean Glycine max, by sequencing and de novo assembly of seven phylogen...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/nbt.2979
更新日期：2014-10-01 00:00:00
abstract：:Field tests of corn co-expressing two new delta-endotoxins from Bacillus thuringiensis (Bt) have demonstrated protection from root damage by western corn rootworm (Diabrotica virgifera virgifera LeConte). The level of protection exceeds that provided by chemical insecticides. In the bacterium, these proteins form crys...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/90282
更新日期：2001-07-01 00:00:00
abstract：:Development of a cell therapy for diabetes would be greatly aided by a renewable supply of human beta-cells. Here we show that pancreatic endoderm derived from human embryonic stem (hES) cells efficiently generates glucose-responsive endocrine cells after implantation into mice. Upon glucose stimulation of the implant...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/nbt1393
更新日期：2008-04-01 00:00:00
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pub_type： 杂志文章
doi：10.1038/s41587-020-0503-6
更新日期：2020-09-01 00:00:00
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pub_type： 杂志文章,评审
doi：10.1038/nbt0207-185
更新日期：2007-02-01 00:00:00
abstract：:Transcriptional silencing, one of the major impediments to gene therapy in humans, is often accompanied by replication during late S-phase. We report that transcriptional silencing and late replication were prevented by DNA sequences that can initiate DNA replication (replicators). When replicators were included in si...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/nbt1202
更新日期：2006-05-01 00:00:00
abstract：:While the use of enzymes as biocatalysts to assist in the industrial manufacture of fine chemicals and pharmaceuticals has enormous potential, application is frequently limited by evolution-led catalyst traits. The advent of designer biocatalysts, produced by informed selection and mutation through recombinant DNA tec...
journal_title：Nature biotechnology
pub_type： 杂志文章,评审
doi：10.1038/nbt0102-37
更新日期：2002-01-01 00:00:00
abstract：:Recent studies have demonstrated the important enzymatic, structural and regulatory roles of RNA in the cell. Here we present a post-transcriptional regulation system in Escherichia coli that uses RNA to both silence and activate gene expression. We inserted a complementary cis sequence directly upstream of the riboso...
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pub_type： 杂志文章
doi：10.1038/nbt986
更新日期：2004-07-01 00:00:00
abstract：:The nonprofit Pew Initiative on Food and Biotechnology is closing, but the need for an independent and neutral body to facilitate dialog on US biotech policy has never been greater. ...
journal_title：Nature biotechnology
pub_type： 社论
doi：10.1038/nbt0207-143
更新日期：2007-02-01 00:00:00
abstract：:We introduce Sailfish, a computational method for quantifying the abundance of previously annotated RNA isoforms from RNA-seq data. Because Sailfish entirely avoids mapping reads, a time-consuming step in all current methods, it provides quantification estimates much faster than do existing approaches (typically 20 ti...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/nbt.2862
更新日期：2014-05-01 00:00:00
abstract：:Despite recent accomplishments, significant challenges remain before cloning can become a routine part of transgenic animal production. ...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/80231
更新日期：2000-10-01 00:00:00
abstract：:The re-use of previously validated designs is critical to the evolution of synthetic biology from a research discipline to an engineering practice. Here we describe the Synthetic Biology Open Language (SBOL), a proposed data standard for exchanging designs within the synthetic biology community. SBOL represents synthe...
journal_title：Nature biotechnology
pub_type： 杂志文章
doi：10.1038/nbt.2891
更新日期：2014-06-01 00:00:00
