Abstract：
:The histamine H(1) receptor (H(1)R) is a key player in acute inflammatory responses. Antihistamines are widely used to relief the symptoms of allergic rhinitis by antagonizing histamine binding to the H(1)R, without possessing intrinsic activity in classical assays such as guinea pig ileum contraction assays or intracellular Ca(2+) mobilization. Overexpression of H(1)R in heterologous cell lines unmasked the capacity of this receptor to signal in a histamine-independent manner. Moreover, a recent screen of therapeutic and reference antagonists on these H(1)R-overexpressing cells revealed that the majority of these drugs are in fact inverse agonists, as they inhibit basal H(1)R activity. In this chapter, we describe several approaches to study H(1)R constitutive signaling that can be used to identify inverse agonists acting at this blockbuster target.
journal_name
Methods Enzymoljournal_title
Methods in enzymologyauthors
Nijmeijer S,Leurs R,Vischer HFdoi
10.1016/B978-0-12-381298-8.00007-1subject
Has Abstractpub_date
2010-01-01 00:00:00pages
127-47eissn
0076-6879issn
1557-7988pii
B978-0-12-381298-8.00007-1journal_volume
484pub_type
杂志文章abstract：:We have shown that Rab34/Rah participates in the promotion of macropinosome formation. Here we describe procedures for the analyses of intracellular localization and some functional properties of Rab34. Rab34 lacks a consensus sequence of the fourth motif for GTP/GDP binding and GTPase activities. Indeed, GTPase assay...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(05)03019-3
更新日期：2005-01-01 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：:Chromatin plays a vital role in regulating cellular processes that occur on the DNA. Modulation of chromatin structure is conducted through interactions with binding factors that direct critical actions such as posttranslational modifications, nucleosome remodeling, and incorporation of histone variants. Specific fact...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-391940-3.00011-1
更新日期：2012-01-01 00:00:00
abstract：:Fertilization by intracytoplasmic sperm injection (ICSI) is a powerful technique that can be used to understand better the biology of fertilization, in addition to a form of assisted reproduction both in humans and in endangered species. Mouse is often the model organism of choice to study mammalian fertilization. The...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(10)76014-6
更新日期：2010-01-01 00:00:00
abstract：:The enzymes involved in mitochondrial oxidative phosphorylation (OXPHOS) are coassembled into higher ordered supercomplexes within the mitochondrial inner membrane. The cytochrome bc(1)-cytochrome c oxidase (COX) supercomplex is formed by the coassociation of the two electron transport chain complexes, the cytochrome ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(08)04411-X
更新日期：2009-01-01 00:00:00
abstract：:In vitro assembly of eukaryotic translation initiation complexes requires purification of ribosomal subunits, eukaryotic initiation factors, and initiator tRNA from natural sources and therefore yields only limited material for functional and structural studies. In this chapter, we describe a robust, affinity chromato...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(07)29005-6
更新日期：2007-01-01 00:00:00
abstract：:Chemokines are essential mediators of leukocyte movement in vivo. In vitro assays of leukocyte migration cannot mimic the complex interactions with other cell types and matrix needed for cells to extravasate and migrate into tissues. Therefore, in vivo strategies to study the effects and potential relevance of chemoki...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2015.09.015
更新日期：2016-01-01 00:00:00
abstract：:Sequence analysis of synthetic peptides using Edman chemistry can be very useful for the elucidation of certain types of synthetic problems, such as residue deletions and the presence of common stable derivatives, and for following the progress of the synthesis itself. However, it can also be a relatively poor techniq...
journal_title：Methods in enzymology
pub_type： 杂志文章,评审
doi：10.1016/s0076-6879(97)89056-8
更新日期：1997-01-01 00:00:00
abstract：:Guanine nucleotide exchange factors (GEFs) are essential regulators of the spatiotemporal conditions of small GTP-binding protein (SMG) activation. Their cellular activities combine the biochemical stimulation of GDP/GTP exchange, which leads to the active conformation of the SMG, to the detection of upstream signals ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(07)38004-X
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：:Two-component signal transduction systems, composed of histidine kinases and response regulators, enable bacteria to sense, respond, and adapt to changes in their internal and external conditions. The importance of these signaling systems is reflected in their widespread distribution and prevalence in the bacterial ki...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(07)23026-5
更新日期：2007-01-01 00:00:00
abstract：:Mitochondria are considered one of the main sources of reactive oxygen species (ROS). The overgeneration of ROS can evoke an intracellular state of oxidative stress, leading to permanent cell damage. Thus, the intracellular accumulation of ROS may not only disrupt the functions of specific tissues and organs but also ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-416618-9.00013-3
更新日期：2014-01-01 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：:Nitric oxide (NO) is a ubiquitous gas with potent biological effects, including vasodilation, neuronal signaling, and antimicrobial activity. NO is a free radical and can readily react with other molecules, in particular, iron centers and oxygen. At physiological concentrations in aqueous solutions, even in the presen...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(08)36007-8
更新日期：2008-01-01 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：:The current treatment of neurological and psychiatric diseases is far beyond being satisfactory. In addition to highly complex disease mechanisms, the blood-brain barrier (BBB) also remains as a challenge by limiting the delivery of the majority of currently available therapeutics to the central nervous system. Severa...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-391860-4.00013-6
更新日期：2012-01-01 00:00:00
abstract：:In vivo genome manipulation through site-directed mutagenesis and chromosome rearrangements has been hindered by the difficulty in achieving high frequencies of targeting and the intensive labor required to create altered genomes that do not contain any heterologous sequence. Here we describe our approach, referred to...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(05)09019-1
更新日期：2006-01-01 00:00:00
abstract：:Electron cryo-microscopy (cryoEM) has advanced dramatically to become a viable tool for high-resolution structural biology research. The ultimate outcome of a cryoEM study is an atomic model of a macromolecule or its complex with interacting partners. This chapter describes a variety of algorithms and software to buil...
journal_title：Methods in enzymology
pub_type： 杂志文章,评审
doi：10.1016/bs.mie.2016.06.003
更新日期：2016-01-01 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：: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...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-801122-5.00009-X
更新日期：2014-01-01 00:00:00
abstract：:Sedimentation velocity experiments measure the transport of molecules in solution under centrifugal force. Here, we describe a method for monitoring the sedimentation of very large biological molecular assemblies using the interference optical systems of the analytical ultracentrifuge. The mass, partial-specific volum...
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：:Human embryonic stem cells hold great promise in furthering our treatment of disease and increasing our understanding of early development. This chapter describes protocols for the derivation and maintenance of human embryonic stem cells. In addition, it summarizes briefly several alternative methods for the culture o...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(06)18005-2
更新日期：2006-01-01 00:00:00
abstract：:Essentially the same steps are required to solve the crystal structure of a serpin as for any other protein: produce and purify protein, grow crystals, collect diffraction data, find estimates of the phase angles, and then refine and validate the structure. For the phasing step, experimental phasing methods involving ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/B978-0-12-385950-1.00004-3
更新日期：2011-01-01 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
pub_type： 杂志文章
doi：10.1016/bs.mie.2019.07.021
更新日期：2020-01-01 00:00:00
abstract：:Vitamin K-dependent carboxylation, an essential posttranslational modification catalyzed by gamma-glutamyl carboxylase, is required for the biological functions of proteins that control blood coagulation, vascular calcification, bone metabolism, and other important physiological processes. Concomitant with carboxylati...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/bs.mie.2016.10.015
更新日期：2017-01-01 00:00:00
abstract：:Numerous aspects of the immune system operate on the basis of complex regulatory networks that are amenable to mathematical and computational modeling. Several modeling frameworks have recently been applied to simulating the immune system, including systems of ordinary differential equations, delay differential equati...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/S0076-6879(09)67004-X
更新日期：2009-01-01 00:00:00
abstract：:Granzyme B (GrB) is the primary molecular mediator of apoptosis by cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells. It is a unique mammalian aspartic acid-cleaving serine protease. On T cell receptor activation, GrB is released from the CTL cytoplasmic granules by exocytosis, enters the target cells and, ...
journal_title：Methods in enzymology
pub_type： 杂志文章
doi：10.1016/s0076-6879(00)22013-2
更新日期：2000-01-01 00:00:00
abstract：:Circadian clocks control daily rhythms in physiology and behavior across all phyla. These rhythms are intrinsic to individual cells that must synchronize to their environment and to each other to anticipate daily events. Recent advances in recording from large numbers of cells for many circadian cycles have enabled re...
journal_title：Methods in enzymology
pub_type： 杂志文章,评审
doi：10.1016/bs.mie.2014.10.042
更新日期：2015-01-01 00:00:00
abstract：:At least two discrete deoxyribonuclease activities can be detected during apoptotic death, one that generates 30- to 500-kilobase pair (kbp) domain-sized fragments and another that mediates internucleosomal DNA degradation. The latter nuclease has been identified as the caspase-activated deoxyribonuclease (CAD)/CPAN, ...
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
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
