Abstract：
:Endosomes are isolated from rat liver using high-speed centrifugation through sucrose density gradients. They are distinguishable from Golgi elements, with which they coisolate, by their capacity to concentrate internalized protein ligands (viz., insulin and epidermal growth factor (EGF)) in receptor-bound intact form. Endosomal signaling to relevant substrates can be readily shown for insulin and EGF receptor tyrosine kinases (RTKs), respectively. Both RTKs undergo dephosphorylation in endosomes. This can be inhibited by the powerful phosphotyrosine phosphatase inhibitors-the peroxovanadium compounds. In vivo administration of these compounds has been shown to activate selectively the endosomal insulin receptor kinase and promote signaling. Taken together, these observations constitute the basis for the signaling endosome hypothesis for which there is now ample evidence. Furthermore, a substantial body of work has documented the importance of endosomal signaling for growth, development, and disease.
journal_name
Methods Enzymoljournal_title
Methods in enzymologyauthors
Posner BI,Bergeron JJdoi
10.1016/B978-0-12-397925-4.00017-1subject
Has Abstractpub_date
2014-01-01 00:00:00pages
293-307eissn
0076-6879issn
1557-7988pii
B978-0-12-397925-4.00017-1journal_volume
535pub_type
杂志文章abstract：:Gene regulatory grids (GRGs) encompass the space of all the possible transcription factor (TF)-target gene interactions that regulate gene expression, with gene regulatory networks (GRNs) representing a temporal and spatial manifestation of a portion of the GRG, essential for the specification of gene expression. Thus...
journal_title：Methods in enzymology
pub_type： 杂志文章
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abstract：:Since the onset of the genomic era more than 1000 bacterial genomes have been sequenced and several fold more are expected to be completed in the near future. These genome sequences supply a wealth of information that can be exploited by statistical methods to gain significant insights into cellular processes. In Volu...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(10)71002-8
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abstract：:Ecological samples of fungal associations pose particular challenges for nucleic acid extraction due to the presence of several genomes. Thorough examination of the samples prior to extraction is important to assess the risks of contamination. If manual separation of symbionts or their axenic cultivation is not feasib...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(05)95004-0
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abstract：:The recalcitrance of the cell wall to enzymatic hydrolysis represents one of the greatest challenges for using biomass to replace the petroleum as a feedstock for fuels and chemicals. Cell walls are complex in architecture and composition, posing a biochemical challenge for the development of efficient enzymes to rele...
journal_title：Methods in enzymology
pub_type： 杂志文章
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更新日期：2012-01-01 00:00:00
abstract：:This chapter reviews basic concepts of nonlinear fluorescence upconversion, a technique whose temporal resolution is essentially limited only by the pulse width of the ultrafast laser. Design aspects for upconversion spectrophotofluorometers are discussed, and a recently developed system is described. We discuss appli...
journal_title：Methods in enzymology
pub_type： 杂志文章,评审
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更新日期：2008-01-01 00:00:00
abstract：:Microbes are now well recognized as major drivers of the biogeochemical cycling that fuels the Earth, and their viruses (phages) are known to be abundant and important in microbial mortality, horizontal gene transfer, and modulating microbial metabolic output. Investigation of environmental phages has been frustrated ...
journal_title：Methods in enzymology
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abstract：:A fundamental concept of phosphorylation-mediated signaling is the precise switching between discrete functional conformations. According to the traditional view, phosphorylation induces a new, active conformation. In this chapter, a series of NMR experiments performed on a response regulator are described that challe...
journal_title：Methods in enzymology
pub_type： 杂志文章
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abstract：:A procedure for an in vitro signaling assay is described for the MAPK and NFkappaB pathways. The method uses a membrane-cleared lysate that contains all the soluble components required for activating these signaling cascades. The pathways can be activated by variety of molecules, including kinases, G-proteins, and E3 ...
journal_title：Methods in enzymology
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abstract：:This chapter outlined the methods used to study intra- and extracellular modifications of apolipoproteins. These and other related studies have shown that several of the apolipoproteins undergo a series of intra- and extracellular modifications as follows: All apolipoproteins studied contain an 18-26 long signal pepti...
journal_title：Methods in enzymology
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journal_title：Methods in enzymology
pub_type： 杂志文章
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更新日期：2012-01-01 00:00:00
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journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(09)68009-5
更新日期：2009-01-01 00:00:00
abstract：:Histone variants and posttranslational modifications (PTMs) are essential for epigenetic regulation of transcriptional expression. Single and/or combinatorial PTMs of histones play important roles in development and disease formation. Mass spectrometry (MS) has been a powerful tool to study histone variants and PTMs, ...
journal_title：Methods in enzymology
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更新日期：2012-01-01 00:00:00
abstract：:Immunogenic cell death (ICD), induced by certain anticancer chemotherapeutics, leads to the emission of danger associated molecular patterns (DAMP) by cancer cells, which facilitates the attraction, activation and maturation of dendritic cells (DC) as well as the subsequent priming of effector T cells. In this context...
journal_title：Methods in enzymology
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abstract：:Antibodies are excellent binding proteins that have found numerous applications in biological research, biotechnology, and medicine. Characterization of their ligand binding properties has long been, and continues to be, the focus of many researchers. Antibodies are also perfect test systems which can be used for the ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-405539-1.00005-1
更新日期：2013-01-01 00:00:00
abstract：:The Microsealed Drug Delivery (MDD) system was developed to achieve several objectives: to maximize the bioavailability of a therapeutic agent in a target tissue; to minimize the adverse side effects of a therapeutic agent or its metabolites; to optimize the onset, rate, and duration of drug delivery; to maintain the...
journal_title：Methods in enzymology
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journal_title：Methods in enzymology
pub_type： 杂志文章
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更新日期：2013-01-01 00:00:00
abstract：:Focal radiation therapy has the potential to generate systemic tumor-targeting immune responses so potent as to eradicate anatomically distant, non-irradiated malignant lesions, a phenomenon commonly referred to as "the abscopal response." In cancer patients, bona fide abscopal responses are rare, although the recent ...
journal_title：Methods in enzymology
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doi：10.1016/bs.mie.2019.04.014
更新日期：2020-01-01 00:00:00
abstract：:Mdm2 is a negative regulator of p53 activity and functions as an E3 ubiquitin ligase of p53. Inhibition of mdm2 E3 ligase activity will block ubiquitination and subsequent proteasome-mediated degradation of p53, resulting in the stabilization of p53 protein that could lead to the restoration of its tumor-suppressor ac...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(05)99046-0
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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：: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 diver...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(10)70027-6
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abstract：:Extracellular thiol/disulfide redox environments are highly regulated in healthy individuals and become oxidized in disease. This oxidation affects the function of cell surface receptors, ion channels, and structural proteins. Downstream signaling due to changes in extracellular redox potential can be studied using a ...
journal_title：Methods in enzymology
pub_type： 杂志文章
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journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2018.04.003
更新日期：2018-01-01 00:00:00
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journal_title：Methods in enzymology
pub_type： 杂志文章
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更新日期：2013-01-01 00:00:00
abstract：:The protein kinase C (PKC) family of isoenzymes may be a crucial player in transducing H2O2-induced signaling in a wide variety of physiological and pathophysiological processes. PKCs contain unique structural features that make them highly susceptible to oxidative modification. Depending on the site of oxidation and ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-405881-1.00005-7
更新日期：2013-01-01 00:00:00
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journal_title：Methods in enzymology
pub_type： 杂志文章,评审
doi：10.1016/B978-0-12-391857-4.00004-5
更新日期：2012-01-01 00:00:00
abstract：:Intrinsically disordered proteins (IDPs) sample structurally diverse ensembles. Characterizing the underlying distributions of conformations is a key step toward understanding the structural and functional properties of IDPs. One increasingly popular method for obtaining quantitative information on intramolecular dist...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2018.09.030
更新日期：2018-01-01 00:00:00
abstract：:RNA helicases are associated with every aspect of RNA metabolism and function. A diverse range of RNA helicases are encoded by essentially every organism. While RNA helicases alter gene expression, RNA helicase expression is itself regulated, frequently in response to abiotic stress. Photosynthetic cyanobacteria prese...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-396546-2.00018-8
更新日期：2012-01-01 00:00:00
abstract：:A number of different analytical techniques are now available for the isolation of apoA-I, apoA-II, and apoA-IV. The choice of a particular technique is dependent on the instrumentation available, and the quantity of isolated apolipoprotein required. The isolation and characterization of the separate isoforms and the ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/0076-6879(86)28070-2
更新日期：1986-01-01 00:00:00
abstract：:Isothermal Titration Calorimetry (ITC) provides a sensitive and accurate means by which to study the thermodynamics of RNA folding, RNA binding to small molecules, and RNA-protein interactions. The advent of extremely sensitive instrumentation and the increasing availability of ITC in shared facilities have made it in...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(09)68019-8
更新日期：2009-01-01 00:00:00
abstract：:We describe here the methods we have used to generate selective peptide inhibitors and activators of PKC-mediated signaling. These approaches should be applicable to any signaling event that is dependent on protein-protein interaction. Furthermore, targeting downstream enzymes in signal transduction has been notorious...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/s0076-6879(02)45039-2
更新日期：2002-01-01 00:00:00
