In vitro and in vivo assays for the activity of Drosha complex.

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

authors： Storici F,Resnick MA

更新日期：2006-01-01 00:00:00

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 intrac...

journal_title：Methods in enzymology

authors： Nijmeijer S,Leurs R,Vischer HF

更新日期：2010-01-01 00:00:00

abstract：:Tumor cells acquire distinct genetic characteristics as a means to survive and proliferate indefinitely. Changes in the genetic code can also translate in changes at the protein level, therefore creating a distinguishable signature unique for tumor cells, and absent in normal tissues. The presence of discernable moiet...

journal_title：Methods in enzymology

authors： Battaglia S

更新日期：2020-01-01 00:00:00

abstract：:Membrane-bound pyrophosphatases couple the hydrolysis of inorganic pyrophosphate to the pumping of ions (sodium or protons) across a membrane in order to generate an electrochemical gradient. This class of membrane protein is widely conserved across plants, fungi, archaea, and bacteria, but absent in multicellular ani...

journal_title：Methods in enzymology

authors： Harborne SPD,Strauss J,Turku A,Watson MA,Tuma R,Harris SA,Goldman A

更新日期：2018-01-01 00:00:00

abstract：:Constitutively activating mutations of the TSHR are the major cause for nonautoimmune hyperthyroidism, which is based on ligand independent, permanent receptor activation. Several reports have highlighted the difficulties to determine whether a TSHR mutation is constitutively active or not especially for borderline ca...

journal_title：Methods in enzymology

authors： Mueller S,Jaeschke H,Paschke R

更新日期：2010-01-01 00:00:00

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

authors： Yang F,Ouma WZ,Li W,Doseff AI,Grotewold E

更新日期：2016-01-01 00:00:00

abstract：:The Duffy blood group antigen is a serpentine protein with seven transmembrane domains that is not coupled to G-proteins or other known intracellular effectors. In addition to erythrocytes, it is also expressed in endothelial cells and neurons. In recent years the Duffy antigen has received much attention because of i...

journal_title：Methods in enzymology

authors： Rot A,Horuk R

更新日期：2009-01-01 00:00:00

abstract：:Mutations in regulators and effectors of the Rho GTPases underlie various forms of mental retardation (MR). Among them, oligophrenin-1 (OPHN1), which encodes a Rho-GTPase activating protein, was one of the first Rho-linked MR genes identified. Upon characterization of OPHN1 in hippocampal brain slices, we obtained evi...

journal_title：Methods in enzymology

authors： Kasri NN,Govek EE,Van Aelst L

更新日期：2008-01-01 00:00:00

abstract：:The Gene Expression Omnibus (GEO) repository at the National Center for Biotechnology Information archives and freely distributes high-throughput molecular abundance data, predominantly gene expression data generated by DNA microarray technology. The database has a flexible design that can handle diverse styles of bot...

journal_title：Methods in enzymology

authors： Barrett T,Edgar R

更新日期：2006-01-01 00:00:00

abstract：:Cholestatic syndromes are inborn or acquired disorders of bile formation. In recent years, several inherited cholestatic syndromes were characterized at the molecular level: progressive familial intrahepatic cholestasis (PFIC) and benign recurrent intrahepatic cholestasis (BRIC). Both PFIC and BRIC were divided phenot...

journal_title：Methods in enzymology

authors： Kubitz R,Keitel V,Häussinger D

更新日期：2005-01-01 00:00:00

abstract：:Neurofilament biology is important to understanding structural properties of axons, such as establishment of axonal diameter by radial growth. In order to study the function of neurofilaments, a series of genetically modified mice have been generated. Here, we describe a brief history of genetic modifications used to ...

journal_title：Methods in enzymology

authors： Jones MR,Villalón E,Garcia ML

更新日期：2016-01-01 00:00:00

abstract：:The fact that chemical exchange processes occur at rates that cover a broad range and produce readily detectable effects on the spectrum is one of the attractive features of high-resolution NMR. The description of these line shapes in the presence of spin-spin coupling requires the density matrix theory which is rathe...

journal_title：Methods in enzymology

authors： Rao BD

更新日期：1989-01-01 00:00:00

abstract：:RNA folds during transcription in the cell. Compared to most in vitro studies where the focus is generally on Mg(2+)-initiated refolding of fully synthesized transcripts, cotranscriptional RNA folding studies better replicate how RNA folds in a cellular environment. Unique aspects of cotranscriptional folding include ...

journal_title：Methods in enzymology

authors： Wong TN,Pan T

更新日期：2009-01-01 00:00:00

abstract：:Phosphoinositide-3-kinase (PI3K) signaling has been implicated in a panoply of cellular responses including survival, proliferation, protein synthesis, migration, and vesicular trafficking. In addition, alterations in the enzymatic activity of PI3Ks have been involved in the pathogenesis of multiple diseases, ranging ...

journal_title：Methods in enzymology

authors： Ciraolo E,Gulluni F,Hirsch E

更新日期：2014-01-01 00:00:00

abstract：:This chapter deals with the use of functionalized carbon nanotubes (fCNTs) as supports for the development of nanobiocatalytic systems through the immobilization of enzymes. The surface characteristics, properties and production of carbon nanotubes are described, while an analysis in their biological applications is a...

journal_title：Methods in enzymology

authors： Patila M,Chalmpes N,Dounousi E,Stamatis H,Gournis D

更新日期：2020-01-01 00:00:00

abstract：:Hematopoietic stem cells (HSCs) primarily reside in bone marrow, are defined by their ability to maintain blood homeostasis, and replenish themselves through self-renewal. Although HSC purification schemes vary from laboratory to laboratory, the resulting cell populations are similar, if not the same. This chapter wil...

journal_title：Methods in enzymology

authors： Lin KK,Goodell MA

更新日期：2006-01-01 00:00:00

abstract：:gamma-Glutamyltranspeptidase is a heterodimeric glycoprotein that catalyzes the transpeptidation and hydrolysis of the gamma-glutamyl group of glutathione and related compounds. It is known that the enzyme plays a role in the metabolism of glutathione and in salvaging constituents of glutathione. In the adult animal, ...

journal_title：Methods in enzymology

authors： Ikeda Y,Taniguchi N

更新日期：2005-01-01 00:00:00

abstract：:Respiration is widely used for evaluation of the metabolic capabilities or physiological state of the microbial culture. This chapter describes novel approaches for characterization of respiration at a single cell level: (1) flow cytometry-based redox sensing (FCRS) of actively metabolizing microbes; (2) respiration r...

journal_title：Methods in enzymology

authors： Konopka MC,McQuaide S,Ojala DS,Kalyuzhnaya MG,Lidstrom ME

更新日期：2011-01-01 00:00:00

abstract：:Covalent modification of proteins with ubiquitin plays an important role in a wide array of cellular processes (Hershko and Ciechanover, 1998; Pickart, 2004). For this reason an increasing number of investigators in diverse research fields are confronted with the question whether their favorite proteins are ubiquitina...

journal_title：Methods in enzymology

authors： Kaiser P,Tagwerker C

更新日期：2005-01-01 00:00:00

abstract：:The relative structural conservation of "internal glycans" in the cell walls of pathogens suggests that they might as target epitopes less prone to variation and hence with greater potential universality as vaccine targets. Examples of such glycans include the inner core sugars of lipopolysaccharides in Gram-negative ...

journal_title：Methods in enzymology

authors： Lakshminarayanan A,Vijayakrishnan B,Bayley H,Davis BG

更新日期：2017-01-01 00:00:00

abstract：:Antimicrosomal antibodies in inflammatory liver diseases all seem to be directed against members of the cytochrome P450 family of proteins. These autoantigens seem to be genetically polymorphic, the autoantibodies are inhibitory, and the autoepitopes are generally conserved among species. Anti-P450 autoantibodies shar...

journal_title：Methods in enzymology

authors： Manns MP,Johnson EF

更新日期：1991-01-01 00:00:00

abstract：:Microsatellite markers are quickly becoming the molecular marker of choice for studies of parentage and kinship in animals. In this chapter, we review methods and give protocols for screening potential microsatellite markers, as well as protocols for genotyping individuals with useful markers once they have been ident...

journal_title：Methods in enzymology

authors： Webster MS,Reichart L

更新日期：2005-01-01 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

authors： Akutsu H,Cowan CA,Melton D

更新日期：2006-01-01 00:00:00

abstract：:Circadian rhythms generated by cell autonomous biological clocks allow for the appropriate temporal synchronization of physiology and behavior, optimizing the efficiency of biological systems. Circadian oscillators and functions have been uncovered in both central and peripheral tissues. This article describes methodo...

journal_title：Methods in enzymology

authors： Rudic RD,Curtis AM,Cheng Y,FitzGerald G

更新日期：2005-01-01 00:00:00

abstract：:β-arrestins (βarrs) are multifunctional proteins that interact with activated and phosphorylated G protein-coupled receptors (GPCRs) to regulate their signaling and trafficking. Understanding the intricate details of GPCR-βarr interaction continues to be a key research area in the field of GPCR biology. Bimane fluores...

journal_title：Methods in enzymology

authors： Srivastava A,Baidya M,Dwivedi-Agnihotri H,Shukla AK

更新日期：2020-01-01 00:00:00

abstract：:Recent advances in functional genomics have enabled genome-wide genetic studies in mammalian cells. These include the establishment of high-throughput transfection and viral propagation methodologies, the production of large-scale cDNA and siRNA libraries, and the development of sensitive assay detection processes and...

journal_title：Methods in enzymology

authors： Rines DR,Tu B,Miraglia L,Welch GL,Zhang J,Hull MV,Orth AP,Chanda SK

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

authors： Read RJ,Zhou A,Stein PE

更新日期：2011-01-01 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

authors： DiMaio F,Chiu W

更新日期：2016-01-01 00:00:00

abstract：:Live cell microscopy using fluorescent proteins and small fluorescent probes is a well-established and essential tool for cell biology; however, there is a considerable need for noninvasive techniques able to study tissue and cell dynamics without the need to introduce chemical or genetically encoded probes. Coherent ...

journal_title：Methods in enzymology

authors： Pope I,Langbein W,Borri P,Watson P

更新日期：2012-01-01 00:00:00

abstract：:DNA repair complexes play crucial roles in maintaining genome integrity, which is essential for the survival of an organism. The understanding of their modes of action is often obscure due to limited structural knowledge. Structural characterizations of these complexes are often challenging due to a poor protein produ...

journal_title：Methods in enzymology

authors： Sawicka M,Aramayo R,Ayala R,Glyde R,Zhang X

更新日期：2017-01-01 00:00:00