Genetic circuitry modulating notch signals through endosomal trafficking.

abstract：:The study of hormone secretion by anterior pituitary cells is complicated by the presence of multiple cell types. For unambiguous interpretation of data it is necessary to identify the cells from which measurements are made. We have described a reliable experimental approach involving the identification of cultured ce...

journal_title：Methods in enzymology

authors： Croxton TL,Armstrong WM,Ben-Jonathan N

更新日期：1989-01-01 00:00:00

abstract：:Using the expression strategies described here, we have demonstrated a model system whereby the sequential signaling events involved in cell proliferation and subsequent transformation regulated by G alpha 12 can be investigated. The model system presented here can also be used to study the temporal interrelationships...

journal_title：Methods in enzymology

authors： Dermott JM,Dhanasekaran N

更新日期：2002-01-01 00:00:00

abstract：:Guanine nucleotide dissociation inhibitor (GDI) is a central regulator of Rab GTPase family members. GDI recycles Rab proteins from the membrane and sequesters the inactive GDP-bound form of Rab in the cytosol for use in multiple rounds of transport. The balance between the membrane-bound form of Rab and the cytosolic...

journal_title：Methods in enzymology

authors： Chen CY,Sakisaka T,Balch WE

更新日期：2005-01-01 00:00:00

abstract：:Microtubule (MT)-based motor proteins transport many cellular factors to their functionally relevant locations within cells, and defects in transport are linked to human disease. Understanding the mechanism and regulation of this transport process in living cells is difficult because of the complex in vivo environment...

journal_title：Methods in enzymology

authors： Barak P,Rai A,Dubey AK,Rai P,Mallik R

更新日期：2014-01-01 00:00:00

abstract：:The methods described here concern the use specialized glassware to achieve accurate sub-micromolar to low-millimolar dioxygen concentrations specifically for the study of flavoproteins. Since the early 1960s, these methods have been passed through the lineage of flavin enzymologists almost exclusively as an oral trad...

journal_title：Methods in enzymology

authors： Moran GR

更新日期：2019-01-01 00:00:00

abstract：:Peptides corresponding to the calmodulin-binding domain of MLCK and other target enzymes promise to be useful tools in the study of calmodulin action. Not only will they help to elucidate the molecular interactions of calmodulin with specific target enzymes, but they should also prove to be useful calmodulin antagonis...

journal_title：Methods in enzymology

authors： Blumenthal DK,Krebs EG

更新日期：1987-01-01 00:00:00

abstract：:Stable isotope tracers are widely used to quantify metabolic rates, and yet a limited number of studies have considered the impact of analytical error on estimates of flux. For example, when estimating the contribution of de novo lipogenesis, one typically measures a minimum of four isotope ratios, i.e., the precursor...

journal_title：Methods in enzymology

authors： Previs SF,Herath K,Castro-Perez J,Mahsut A,Zhou H,McLaren DG,Shah V,Rohm RJ,Stout SJ,Zhong W,Wang SP,Johns DG,Hubbard BK,Cleary MA,Roddy TP

更新日期：2015-01-01 00:00:00

abstract：:Helical symmetry is commonly used for building macromolecular assemblies. Helical symmetry is naturally present in viruses and cytoskeletal filaments and also occurs during crystallization of isolated proteins, such as Ca-ATPase and the nicotinic acetyl choline receptor. Structure determination of helical assemblies b...

journal_title：Methods in enzymology

authors： Diaz R,Rice WJ,Stokes DL

更新日期：2010-01-01 00:00:00

abstract：:Integrin adhesion receptors mediate cell-cell and cell-substratum adhesion and provide a continuous link for the bidirectional transmission of mechanical force and biochemical signals across the plasma membrane. Integrin-dependent cellular activities such as adhesion, migration, proliferation, and survival rely upon t...

journal_title：Methods in enzymology

authors： Lad Y,Harburger DS,Calderwood DA

更新日期：2007-01-01 00:00:00

abstract：:Nucleic acid aptamers find widespread use as targeting and sensing agents in nature and biotechnology. Their ability to bind an extensive range of molecular targets, including small molecules, proteins, and ions, with high affinity and specificity enables their use in diverse diagnostic, therapeutic, imaging, and gene...

journal_title：Methods in enzymology

authors： Chang AL,McKeague M,Smolke CD

更新日期：2014-01-01 00:00:00

abstract：:Peptides are multidentate chiral ligands capable of coordinating different metal ions. Nowadays, they can be obtained with high yield and purity, thanks to the advances on peptide/protein chemistry as well as in equipment (peptide synthesizers). Based on the identity and length of their amino acid sequences, peptides ...

journal_title：Methods in enzymology

authors： Lima LM,Iranzo O

更新日期：2016-01-01 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：:Benzylisoquinoline alkaloids (BIAs) constitute a diverse class of plant secondary metabolites that includes the opiate analgesics morphine and codeine. Collectively, BIAs exhibit a myriad of pharmacological activities, including antimicrobial, antitussive, antispasmodic, and anticancer properties. Despite 2500 known B...

journal_title：Methods in enzymology

authors： Pyne ME,Narcross L,Fossati E,Bourgeois L,Burton E,Gold ND,Martin VJ

更新日期：2016-01-01 00:00:00

abstract：:Gene manipulation serves the purpose of providing a better understanding of the function of specific genes as well as for developing novel variants of the genes of interest. The generation of knockout genes, the alteration, depletion, or enhancement of a particular gene function through the generation of specific gene...

journal_title：Methods in enzymology

authors： Stuckey S,Storici F

更新日期：2013-01-01 00:00:00

abstract：:Intracellular protein degradation is one of the most precisely regulated processes in living cells. The main component of the degradation machinery is the 20S proteasome present in eukaryotes as well as in prokaryotes. We have developed successful purification protocols for the 20S proteasome in its native state using...

journal_title：Methods in enzymology

authors： Groll M,Huber R

更新日期：2005-01-01 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：:Synthetic hydrogen sulfide (H2S) donors are useful research tools as well as potential therapeutic agents. In this chapter, we report the detailed protocols for the synthesis and evaluation of a series of phosphorodithioate-based H2S donors. Fluorescence assays were used to determine H2S release from the donors in bot...

journal_title：Methods in enzymology

authors： Park CM,Xian M

更新日期：2015-01-01 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：:Mitochondrial RNAs in the myxomycete Physarum polycephalum differ from the templates from which they are transcribed in defined ways. Most transcripts contain nucleotides that are not present in their respective genes. These "extra" nucleotides are added during RNA synthesis by an unknown mechanism. Other differences ...

journal_title：Methods in enzymology

authors： Byrne EM,Visomirski-Robic L,Cheng YW,Rhee AC,Gott JM

更新日期：2007-01-01 00:00:00

abstract：:Obtaining patient-specific information through the quantification of small molecules and proteins in bodily fluids is essential for personalized therapies. Point-of-care (POC) diagnostic devices hold the promise of delivering such benefit to a wide range of patients. However, there is a lack of enabling technology, as...

journal_title：Methods in enzymology

authors： Yu Q,Griss R,Schena A,Johnsson K

更新日期：2017-01-01 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：:Astrocytes are the most abundant cell type in the brain and are a crucial part of solving its mysteries. Originally assumed to be passive supporting cells, astrocyte's functions are now recognized to include active modulation and information processing at the neural synapse. The full extent of the astrocyte contributi...

journal_title：Methods in enzymology

authors： Preston AN,Cervasio DA,Laughlin ST

更新日期：2019-01-01 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

authors： Marin-Sanguino A,Gupta SK,Voit EO,Vera J

更新日期：2011-01-01 00:00:00

abstract：:Liquid-liquid phase separation (LLPS) has been known to drive formation of biomolecular compartments, which can encapsulate RNA and proteins among other cosolutes. Such compartments, which lack a lipid membrane, have been implicated in origins of life scenarios as they can easily uptake and concentrate biomolecules, s...

journal_title：Methods in enzymology

authors： Poudyal RR,Meyer MO,Bevilacqua PC

更新日期：2021-01-01 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

authors： Reigstad LJ,Bartossek R,Schleper C

更新日期：2011-01-01 00:00:00

abstract：:Cell wall precursors that have been modified at their peptide moiety were incorporated into the living bacterial cell wall. Using chemically synthesized bacterial cell wall precursors, a variety of compounds could be attached to the bacterial surface. Escherichia coli took the modified precursors into the cell wall af...

journal_title：Methods in enzymology

authors： Sadamoto R,Niikura K,Monde K,Nishimura S

更新日期：2003-01-01 00:00:00

abstract：:Dicer is a multidomain ribonuclease that processes double-stranded RNAs (dsRNAs) to 21-nt small interfering RNAs (siRNAs) during RNA interference and excises microRNAs (miRNAs) from precursor hairpins. PAZ and PIWI domain (PPD) proteins, also involved in RNAi and miRNA function, are the best-characterized proteins kno...

journal_title：Methods in enzymology

authors： Kolb FA,Zhang H,Jaronczyk K,Tahbaz N,Hobman TC,Filipowicz W

更新日期：2005-01-01 00:00:00

abstract：:Undiluted cytoplasmic extract prepared from unfertilized Xenopus laevis eggs by low-speed centrifugation (CSF extracts) is useful for reconstitution of egg microtubule dynamics and meiosis-II spindle organization, but it suffers limitations for biochemical analysis due to abundant particulates. Here, we describe prepa...

journal_title：Methods in enzymology

authors： Groen AC,Ngyuen PA,Field CM,Ishihara K,Mitchison TJ

更新日期：2014-01-01 00:00:00

abstract：:Histone acetylation is a dynamic epigenetic modification that functions in the regulation of DNA-templated reactions, such as transcription. This lysine modification is reversibly controlled by histone (lysine) acetyltransferases and deacetylases. Here, we present methods employing isotopic labeling and mass spectrome...

journal_title：Methods in enzymology

authors： Fan J,Baeza J,Denu JM

更新日期：2016-01-01 00:00:00

abstract：:High content, phenotypic screens offer a powerful approach to systems biology at the cellular level. The approach employs cells carrying fluorescently labeled molecules or organelles in 384- or 1536-well microplates, and an automated confocal screening microscope for capturing images from each well. Although some spec...

journal_title：Methods in enzymology

authors： Kepiro M,Varkuti BH,Davis RL

更新日期：2018-01-01 00:00:00