Abstract：
:Approximately one-third of all newly translated polypeptides interact with the endoplasmic reticulum (ER), an event that is essential to target these nascent proteins to distinct compartments within the cell or to the extracellular milieu. Thus, the ER houses molecular chaperones that augment the folding of this diverse group of macromolecules. The ER also houses the enzymes that catalyze a multitude of posttranslational modifications. If, however, proteins misfold or are improperly modified in the ER they are proteolyzed via a process known as ER-associated degradation (ERAD). During ERAD, substrates are selected by molecular chaperones and chaperone-like proteins. They are then delivered to the cytoplasmic proteasome and hydrolyzed. In most cases, delivery and proteasome-targeting require the covalent attachment of ubiquitin. The discovery and underlying mechanisms of the ERAD pathway have been aided by the development of in vitro assays that employ components derived from the yeast, Saccharomyces cerevisiae. These assays recapitulate the selection of ERAD substrates, the "retrotranslocation" of selected polypeptides from the ER into the cytoplasm, and the proteasome-mediated degradation of the substrate. The ubiquitination of integral membrane ERAD substrates has also been reconstituted.
journal_name
Methods Enzymoljournal_title
Methods in enzymologyauthors
Brodsky JLdoi
10.1016/S0076-6879(10)70027-6subject
Has Abstractpub_date
2010-01-01 00:00:00pages
661-79eissn
0076-6879issn
1557-7988pii
S0076-6879(10)70027-6journal_volume
470pub_type
杂志文章abstract：:Protein kinase C (PKC), a family of serine/threonine kinases, plays an important role in apoptosis. Several members of the PKC family act as substrates for caspases. In addition, PKCs can also regulate caspase activation and cell death by apoptosis. The cleavage of PKCs separates the regulatory domain from the catalyt...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(08)01608-X
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abstract：:A central goal of biology is to understand how transcription factors target and regulate specific genes and networks to control cell fate and function. An equally important goal of synthetic biology, chemical biology, and personalized medicine is to devise molecules that can regulate genes and networks in a programmab...
journal_title：Methods in enzymology
pub_type： 杂志文章
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更新日期：2011-01-01 00:00:00
abstract：:Optogenetic systems enable unmatched precision for controlling molecular biological processes but require the use of specialized electrical and optical hardware. In an effort to make working with optogenetic systems more accessible, we recently described an open-source hardware platform called the Light Plate Apparatu...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2019.04.018
更新日期：2019-01-01 00:00:00
abstract：:Complex I is the major entry point in the bacterial and mitochondrial respiratory chain. Structural knowledge of the enzyme is still limited because of its large size and complicated architecture. Only the structure of the hydrophilic domain of a bacterial Complex I has been solved to high resolution by X-ray crystall...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(08)04401-7
更新日期：2009-01-01 00:00:00
abstract：:Single-cell analysis of cellular contents by highly sensitive analytical instruments is known as chemical cytometry. A chemical cytometer typically samples one cell at a time, quantifies the cellular contents of interest, and then processes and reports that data. Automation adds the potential to perform this entire se...
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pub_type： 杂志文章,评审
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journal_title：Methods in enzymology
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journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(05)03049-1
更新日期：2005-01-01 00:00:00
abstract：:Specific recognition of mRNA 5' cap by eukaryotic initiation factor eIF4E is a rate-limiting step in the translation initiation. Structural determination of the eIF4E-cap complexes, as well as complexes of eIF4E with other proteins regulating its activity, requires complementary experiments that allow for energetic an...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(07)30009-8
更新日期：2007-01-01 00:00:00
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journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(06)22019-6
更新日期：2007-01-01 00:00:00
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journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-801185-0.00008-8
更新日期：2014-01-01 00:00:00
abstract：:After cell lysis, the most often used second step in a protein purification procedure is some sort of a rapid, bulk precipitation step. This is commonly accomplished by altering the solvent conditions and taking advantage of the changes in solubility of your protein of interest relative to those of many of the other p...
journal_title：Methods in enzymology
pub_type： 杂志文章,评审
doi：10.1016/S0076-6879(09)63020-2
更新日期：2009-01-01 00:00:00
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journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2019.04.014
更新日期：2020-01-01 00:00:00
abstract：:In actively growing cells, the rate of translation initiation is relatively rapid. As a result, multiple ribosomes simultaneously engaged in translation become spaced along single mRNA molecules. These structures, termed polysomes, can be separated from ribosomal subunits and single ribosomes because they migrate fast...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-420037-1.00008-7
更新日期：2013-01-01 00:00:00
abstract：:We describe a method for molecular confinement and single-fluorophore sensitive measurement in aqueous nanodroplets in oil. The sequestration of individual molecules in droplets has become a useful tool in genomics and molecular evolution. Similarly, the use of single fluorophores, or pairs of fluorophores, to study b...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(10)72015-2
更新日期：2010-01-01 00:00:00
abstract：:Pathogen-host interactions are mediated in part by secreted microbial proteins capable of exploiting host cells for their survival. Several of these manipulations involve, but are not limited to, suppression of defense responses, alterations in host vesicular trafficking, and manipulation of gene expression. The deliv...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-397925-4.00007-9
更新日期：2014-01-01 00:00:00
abstract：:In the near future, computational tools and methods based on the mathematical modeling of biomedically relevant networks and pathways will be necessary for the design of therapeutic strategies that fight complex multifactorial diseases. Beyond the use of pharmacokinetic and pharmacodynamic approaches, we propose here ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-381270-4.00011-1
更新日期：2011-01-01 00:00:00
abstract：:MicroRNA (miRNA) genes are transcribed into long primary transcripts (pri-miRNAs) that get processed into mature miRNAs of about 22 nt in length by two different ribonuclease (RNase) III enzymes, Drosha and Dicer. Various experimental protocols have been developed and modified for genetic and biochemical analyses for ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(07)27005-3
更新日期：2007-01-01 00:00:00
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journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/s0076-6879(00)22003-x
更新日期：2000-01-01 00:00:00
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journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-417158-9.00007-8
更新日期：2014-01-01 00:00:00
abstract：:Technical innovations in structural probing have drastically advanced the field of RNA structure analysis. These advances have led to parallel approaches developed in separate labs for analyzing RNA structure and dynamics. With the wealth of methodologies available, it can be difficult to determine which is best suite...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2019.04.002
更新日期：2019-01-01 00:00:00
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journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2015.05.027
更新日期：2015-01-01 00:00:00
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journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2016.11.009
更新日期：2017-01-01 00:00:00
abstract：:Osmoregulation is the active control of the cellular water balance and encompasses homeostatic mechanisms crucial for life. The osmoregulatory system in the yeast Saccharomyces cerevisiae is particularly well understood. Key to yeast osmoregulation is the production and accumulation of the compatible solute glycerol, ...
journal_title：Methods in enzymology
pub_type： 杂志文章,评审
doi：10.1016/S0076-6879(07)28002-4
更新日期：2007-01-01 00:00:00
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journal_title：Methods in enzymology
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更新日期：2010-01-01 00:00:00
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journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2015.05.007
更新日期：2015-01-01 00:00:00
abstract：:We have developed an array-based whole-genome genotyping (WGG) assay (Infinium) using our BeadChip platform that effectively enables unlimited multiplexing and unconstrained single nucleotide polymorphism (SNP) selection. A single tube whole-genome amplification reaction is used to amplify the genome, and loci of inte...
journal_title：Methods in enzymology
pub_type： 杂志文章,评审
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更新日期：2006-01-01 00:00:00
abstract：:Reference standard materials (RSMs) exist for a variety of biologics including vaccines but are not readily available for gene therapy vectors. To date, a recombinant adeno-associated virus serotype 2 RSM (rAAV2 RSM) has been produced and characterized and was made available to the scientific community in 2010. In add...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-386509-0.00015-6
更新日期：2012-01-01 00:00:00
abstract：:Rab38 is a low-molecular-weight G-protein highly expressed in melanocytes of the skin and alveolar type II cells in the lung. A point mutation in the postulated GTP/GDP-interacting domain of Rab38 has been identified as the genetic lesion responsible for oculocutaneous albinism (OCA) in chocolate (cht) mice. Another p...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(07)38014-2
更新日期：2008-01-01 00:00:00
abstract：:A synthetic ligand regulable system for gene transfer and expression has been developed in our laboratory based on mechanistic studies of steriod hormone receptor and transcriptional regulation. This gene switch system possesses most of the important features that are required for application of the system in biologic...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/s0076-6879(02)46076-4
更新日期：2002-01-01 00:00:00
