Abstract：
:Signal transduction by vascular endothelial growth factors (VEGFs) through their cognate VEGF receptor tyrosine kinases follows the consensus scheme for receptor tyrosine kinases. Thus, binding of ligand induces receptor dimerization and activation of the tyrosine kinase through transphosphorylation between receptor molecules, leading to initiation of intracellular signal transduction pathways. Certain signal transduction pathways are shared with most, if not all, receptor tyrosine kinases, whereas some may be unique (e.g., transduced only by VEGF receptors). Indications that such unique signaling pathways may be discerned only when VEGF receptors are expressed in their proper context (i.e., in endothelial cells of microcapillary origin). In this chapter, we describe a number of methods for the study of signal transduction in endothelial cells. We describe how to isolate and examine endothelial cell lines. We also describe the embryoid body model representing vasculogenesis and angiogenesis, the procedure for subcutaneous Matrigel plugs, and, finally, how to construct gene-targeted mouse models. We emphasize the need for validation of in vitro data in more complex models, where endothelial cells reside in their proper three-dimensional context.
journal_name
Methods Enzymoljournal_title
Methods in enzymologyauthors
Li X,Claesson-Welsh L,Shibuya Mdoi
10.1016/S0076-6879(08)02013-2subject
Has Abstractpub_date
2008-01-01 00:00:00pages
261-84eissn
0076-6879issn
1557-7988pii
S0076-6879(08)02013-2journal_volume
443pub_type
杂志文章abstract：:Considering that HIV-1 accumulates and replicates actively within lymphoid tissues, any strategy that will decrease viral stores in these tissues might be beneficial to the infected host. Follicular dendritic cells (FDC), B lymphocytes, antigen-presenting cells like macrophages, and activated CD4(+) T cells are abunda...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(05)91019-7
更新日期：2005-01-01 00:00:00
abstract：:Delivery of drugs and molecules to eukaryote animal and plant cells can be enhanced when mediated by mesoporous silica nanoparticle (MSN) materials. This chapter describes, in detail, the synthesis of several different MSN materials used for the transport of various hydrophilic and hydrophobic drug molecules, neurotra...
journal_title：Methods in enzymology
pub_type： 杂志文章,评审
doi：10.1016/B978-0-12-391860-4.00003-3
更新日期：2012-01-01 00:00:00
abstract：:Cells belonging to the monocyte/macrophage lineage are in general highly resistant to peroxynitrite. Resistance is not dependent on the scavenging of peroxynitrite itself, or of other secondary reactive species, but is rather associated with the prompt activation of a survival signaling leading to the prevention of to...
journal_title：Methods in enzymology
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doi：10.1016/S0076-6879(08)01205-6
更新日期：2008-01-01 00:00:00
abstract：:Given recent efforts to determine the sequence information on thousands of genes in the human genome, the current challenge is to identify the functions of these genes, including those encoding the regulator of G-protein signaling protein gene superfamily, and to establish their roles in particular signaling pathways ...
journal_title：Methods in enzymology
pub_type： 杂志文章,评审
doi：10.1016/S0076-6879(04)89015-3
更新日期：2004-01-01 00:00:00
abstract：:Atomic force microscopy (AFM) enables direct visualization of single-protein molecules in liquids at submolecular resolution. High-speed AFM further makes it possible to visualize dynamic biomolecular processes at subsecond resolution. However, dynamic imaging of biomolecular processes imposes various requirements on ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(10)75020-5
更新日期：2010-01-01 00:00:00
abstract：:RNA and DNA carry out diverse functions in biology including catalysis, splicing, gene regulation, and storage of genetic information. Interest has grown in understanding how nucleic acids perform such sophisticated functions given their limited molecular repertoire. RNA can fold into diverse shapes that often perturb...
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doi：10.1016/B978-0-12-801122-5.00009-X
更新日期：2014-01-01 00:00:00
abstract：:In structural proteomics, it is necessary to efficiently screen in a high-throughput manner for the presence of stable structures in proteins that can be subjected to subsequent structure determination by X-ray or NMR spectroscopy. Here we illustrate that the (1)H chemical distribution in a protein as detected by (1)H...
journal_title：Methods in enzymology
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doi：10.1016/S0076-6879(05)94006-8
更新日期：2005-01-01 00:00:00
abstract：:The mapping and characterization of the vasculature using proteomics offers the opportunity to better understand the steps and molecular mechanisms involved in vascular development, and angiogenesis in particular. Proteomics has many key advantages over genomics, especially in directly determining protein expression. ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(08)03008-5
更新日期：2008-01-01 00:00:00
abstract：:Reviewing the circumstances that have led to the first monoclonal antibody against the disease-associated form of PrP, we consider the availability of PrP knockout mice and recombinant PrP, as well as a reliable conformational screening protocol as being important prerequisites for a successful immunization approach. ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/s0076-6879(99)09010-2
更新日期：1999-01-01 00:00:00
abstract：:The RNA-guided, sequence-specific endonuclease Cas9 has been widely adopted as genome engineering tool due to its efficiency and ease of use. Derived from the microbial CRISPR (clustered regularly interspaced short palindromic repeat) type II adaptive immune system, Cas9 has now been successfully engineered for genome...
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：: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：:The existence of an accurate force field (FF) model that reproduces the free-energy landscape is a key prerequisite for the simulation of biomineralization. Here, the stages in the development of such a model are discussed including the quality of the water model, the thermodynamics of polymorphism, and the free energ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-416617-2.00001-1
更新日期：2013-01-01 00:00:00
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
doi：10.1016/B978-0-12-385075-1.00001-9
更新日期：2011-01-01 00:00:00
abstract：:Aliphatic carbon-halogen (C-X) bonds are prevalent in modern pharmaceuticals and bioactive natural products. Three distinct chemical strategies are known in Nature to generate these structural motifs. The first is via the nucleophilic substitution at a prefunctionalized electrophilic carbon center with a halide anion ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2018.02.015
更新日期：2018-01-01 00:00:00
abstract：:ADP-ribosylation is a protein post-translational modification that is critically involved in a wide array of biological processes connected to cell stress responses. Enzymes known as poly-ADP-ribose polymerases (PARPs) catalyze the addition of the ADP-ribose units to amino acids with various side chain chemistries. In...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2019.06.024
更新日期：2019-01-01 00:00:00
abstract：:Growing appreciation for the biogeochemical significance of uncultured microorganisms is changing the focus of environmental microbiology. Techniques designed to investigate microbial metabolism in situ are increasingly popular, from mRNA-targeted fluorescence in situ hybridization (FISH) to the "-omics" revolution, i...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-381294-0.00012-2
更新日期：2011-01-01 00:00:00
abstract：:Acidic organelles and vesicles, such as endosomes, lysosomes, autophagosomes, trans-Golgi network, and synaptic vesicles, are known to play important roles in a broad range of cellular events. To facilitate studying these multifunctional systems, we describe here an acid-brightening fluorescent protein (abFP), which f...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2020.04.013
更新日期：2020-01-01 00:00:00
abstract：:Metagenomics has become an important tool for the characterization of microorganisms, as it is independent of their enrichment or cultivation in the laboratory. Its application has led to the discovery of metabolisms from widespread, yet uncharacterized organisms such as the ammonia-oxidizing archaea. Different approa...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-386489-5.00013-0
更新日期：2011-01-01 00:00:00
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...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-397925-4.00017-1
更新日期：2014-01-01 00:00:00
abstract：:The nitrogen (N) cycle is the only global biogeochemical cycle that is driven by biological functions involving the interaction of many microorganisms. The N cycle has evolved over geological time and its interaction with the oxygen cycle has had profound effects on the evolution and timing of Earth's atmosphere oxyge...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-381294-0.00022-5
更新日期：2011-01-01 00:00:00
abstract：:Peptide-based strategies represent transformative approaches to fabricate functional inorganic materials under sustainable conditions by modeling the methods exploited in biology. In general, peptides with inorganic affinity and specificity have been isolated from organisms and through biocombinatorial selection techn...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2016.05.010
更新日期：2016-01-01 00:00:00
abstract：:Omics approaches have become popular in biology as powerful discovery tools, and currently gain in interest for diagnostic applications. Establishing the accurate genome sequence of any organism is easy, but the outcome of its annotation by means of automatic pipelines remains imprecise. Some protein-encoding genes ma...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2016.09.019
更新日期：2017-01-01 00:00:00
abstract：:The remarkable success of cancer immunotherapies, especially the checkpoint blocking antibodies, in a subset of patients has reinvigorated the study of tumor-immune crosstalk and its role in heterogeneity of response. High-throughput sequencing and imaging technologies can help recapitulate various aspects of the tumo...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2020.01.001
更新日期：2020-01-01 00:00:00
abstract：:The ability to efficiently modulate autophagy activity is paramount in the study of the field. Conventional broad-range autophagy inhibitors and genetic manipulation using RNA interference (RNAi), although widely used in autophagy research, are often limited in specificity or efficacy. In this chapter, we address the ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2016.09.076
更新日期：2017-01-01 00:00:00
abstract：:The combined use of rotating-frame relaxation methods, temperature-dependent measurements of line shapes and magnetization transfer experiments allows in favorable cases the examination in some detail of exchange processes that occur on the millisecond time scale. It is possible to determine not only the rate constant...
journal_title：Methods in enzymology
pub_type： 杂志文章,评审
doi：10.1016/s0076-6879(94)39023-1
更新日期：1994-01-01 00:00:00
abstract：:Unfolded protein response (UPR) is a major reaction to intrinsic stress of eukaryotic organisms and is also related to environmental stress reactions. Among yeasts, stress regulation has mainly been investigated in Saccharomyces cerevisiae, while other species with biotechnological or medical interest are less well un...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-385116-1.00010-8
更新日期：2011-01-01 00:00:00
abstract：:Membrane fission, which divides membrane surfaces into separate compartments, is essential to diverse cellular processes including membrane trafficking and cell division. Quantitative assays are needed to elucidate the physical mechanisms by which proteins drive membrane fission. Toward this goal, several experimental...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2018.08.014
更新日期：2018-01-01 00:00:00
abstract：:Flavoenzymes comprise a large class of proteins that carry out a diverse range of important redox chemistry. Although X-ray crystal structures of many flavoenzymes have been determined, there are still unresolved questions regarding the actual oxidation state of the flavin cofactors in these structures due to photored...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2019.12.004
更新日期：2020-01-01 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
doi：10.1016/S0076-6879(08)03408-3
更新日期：2008-01-01 00:00:00
abstract：:Since the early days of cell volume regulation research, the role of actin cytoskeleton organization and rearrangement has attracted specific interest. Rapid modifications in actin dynamics and architecture have been described. They were shown to regulate cell volume changes, as well as regulatory volume decrease in a...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(07)28012-7
更新日期：2007-01-01 00:00:00
