Neural stem cells and their manipulation.

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：:The Arc (anoxic redox control) two-component system (TCS) is a complex signal transduction system that plays an important role in regulating energy metabolism at the level of transcription in bacteria. This system comprises the ArcB protein, a hybrid membrane-associated sensor kinase, and the ArcA protein, a typical r...

journal_title：Methods in enzymology

authors： Alvarez AF,Georgellis D

更新日期：2010-01-01 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：:Regulator of G-protein signaling (RGS) domains bind directly to GTP-bound Galpha subunits and accelerate their intrinsic GTPase activity by up to several thousandfold. The selectivity of RGS proteins for individual Galpha subunits has been illustrated. Thus, the expression of RGS proteins can be used to inhibit signal...

journal_title：Methods in enzymology

authors： Hains MD,Siderovski DP,Harden TK

更新日期：2004-01-01 00:00:00

abstract：:There are estimated to be >300,000 plant species, producing >200,000 metabolites. Many of these metabolites are restricted to specific plant lineages and are referred to as "specialized" metabolites. These serve varied functions in plants including defense against biotic and abiotic stresses, plant-plant and plant-mic...

journal_title：Methods in enzymology

authors： Fan P,Moghe GD,Last RL

更新日期：2016-01-01 00:00:00

abstract：:Vertebrate odorant-binding proteins (OBPs) are small soluble proteins abundantly secreted in the olfactory mucus of many animal species, including humans. Vertebrate OBPs reversibly bind odorant molecules with micromolar range affinities. Although their physiological role is not clearly understood, OBPs are proposed t...

journal_title：Methods in enzymology

authors： Brulé M,Glaz M,Belloir C,Poirier N,Moitrier L,Neiers F,Briand L

更新日期：2020-01-01 00:00:00

abstract：:The aim of this chapter is to discuss the basic principles and reasoning behind elementary flux mode analysis (EFM analysis)--an important tool for the analysis of metabolic networks. We begin with a short introduction into metabolic pathway analysis and subsequently outline in detail fundamentals of EFM analysis by w...

journal_title：Methods in enzymology

authors： Schäuble S,Schuster S,Kaleta C

更新日期：2011-01-01 00:00:00

abstract：:Glass microfluidic devices have been fabricated to monitor the secretion of glycerol or fatty acids from cultured murine 3T3-L1 adipocytes. In the current studies, adipocytes are perfused in a reversibly sealed cell chamber, and secreted products are analyzed by enzyme assay on either a single- or dual-chip device. Th...

journal_title：Methods in enzymology

authors： Dugan CE,Kennedy RT

更新日期：2014-01-01 00:00:00

abstract：:Research on circadian biology over the past decade has paid increasing attention to the photoreceptor mechanisms that align the molecular clock to the 24-h light/dark cycle, and some of the results to emerge are surprising. For example, the rods and cones within the mammalian eye are not required for entrainment. A po...

journal_title：Methods in enzymology

authors： Peirson SN,Thompson S,Hankins MW,Foster RG

更新日期：2005-01-01 00:00:00

abstract：:Liposomes show great promise as intravenous drug delivery vehicles, but it is often difficult to combine stability in the circulation with rapid, targeted release at the site of interest. Targeting to specific tissues requires developing highly specific ligands with strong affinities to receptors overexpressed on dise...

journal_title：Methods in enzymology

authors： Wu G,Mikhailovsky A,Khant HA,Zasadzinski JA

更新日期：2009-01-01 00:00:00

abstract：:Measurement of catalysis by MnSOD using direct observation of the UV absorbance of superoxide allows determination of steady-state catalytic constants. Stabilizing superoxide in aprotic solvents such as dimethyl sulfoxide permits the use of stopped-flow spectrophotometry, although significant information is lost in th...

journal_title：Methods in enzymology

authors： Silverman DN,Nick HS

更新日期：2002-01-01 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：:The circadian clock exists to synchronize inner physiology with the external world, allowing life to anticipate and adapt to the continual changes that occur in an organism's environment. The clock architecture is highly conserved, present in almost all major branches of life. Within eukaryotes, the filamentous fungus...

journal_title：Methods in enzymology

authors： Hurley J,Loros JJ,Dunlap JC

更新日期：2015-01-01 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：:Enzymatic processes have great potential for applications in industrial biocatalysis, biosensing, and biomedical engineering. However, poor stability and difficulty in the reuse of enzymes still create challenges for their use in catalysis and other applications. In recent years, enzyme-incorporated metal-organic fram...

journal_title：Methods in enzymology

authors： Hou M,Ge J

更新日期：2017-01-01 00:00:00

abstract：:Of all arginine catabolic enzymes, the arginases and nitric oxide (NO) synthases are the ones that are of greatest interest to many investigators. Mammalian arginases catalyze the hydrolysis of arginine to ornithine and urea and are composed of two distinct isozymes: arginase I, located within the cytosol, and arginas...

journal_title：Methods in enzymology

authors： Kepka-Lenhart D,Ash DE,Morris SM

更新日期：2008-01-01 00:00:00

abstract：:Death receptors are a subgroup of the tumor necrosis factor receptor superfamily (TNFRSF) and mediate activation of what is widely known as the "extrinsic" apoptosis pathway. TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) is one of the most recent death receptor ligands identified. The TRAIL receptor ...

journal_title：Methods in enzymology

authors： Harper N,MacFarlane M

更新日期：2008-01-01 00:00:00

abstract：:Protein kinase C (PKC), a family of serine/threonine kinases, plays an important role in apoptosis. Several members of the PKC family act as substrates for caspases. In addition, PKCs can also regulate caspase activation and cell death by apoptosis. The cleavage of PKCs separates the regulatory domain from the catalyt...

journal_title：Methods in enzymology

authors： Basu A,Persaud SD,Sivaprasad U

更新日期：2008-01-01 00:00:00

abstract：:The growth and survival of cancer cells are often driven by constitutive activity in the mitogen-activated protein kinase (MAPK) and phospho-inositide 3-kinase (PI3K)/AKT signaling pathways. Activity in these signal transduction cascades is known to contribute to the uncontrolled growth and resistance to apoptosis tha...

journal_title：Methods in enzymology

authors： Paraiso KH,Van Der Kooi K,Messina JL,Smalley KS

更新日期：2010-01-01 00:00:00

abstract：:Nanotechnology offers a new platform for therapeutic delivery of antiretrovirals to the central nervous system (CNS) where human immunodeficiency virus (HIV-1) is sequestered in patients with HIV-1-associated neurological disorders (HAND). HAND is a spectrum of neurocognitive disorders that continue to persist in HIV-...

journal_title：Methods in enzymology

authors： Mahajan SD,Law WC,Aalinkeel R,Reynolds J,Nair BB,Yong KT,Roy I,Prasad PN,Schwartz SA

更新日期：2012-01-01 00:00:00

abstract：:Nonnative protein aggregation has been classically treated as an amorphous process occurring by colloidal coagulation kinetics and proceeding to an essentially irreversible endpoint often ascribed to a chaotic tangle of unfolded chains. However, some nonnative aggregates, particularly amyloid fibrils, exhibit ordered ...

journal_title：Methods in enzymology

authors： O'Nuallain B,Thakur AK,Williams AD,Bhattacharyya AM,Chen S,Thiagarajan G,Wetzel R

更新日期：2006-01-01 00:00:00

abstract：:The identification and isolation of the soluble catalytic domains of adenylyl cyclase have provided investigators with useful reagents for the study of these enzymes. They have permitted detailed mechanistic investigation of the actions of forskolin, Gs alpha, and the inhibitory G protein, Gi alpha. They have served a...

journal_title：Methods in enzymology

authors： Hatley ME,Gilman AG,Sunahara RK

更新日期：2002-01-01 00:00:00

abstract：:Small GTPases of the Ras and Rho families are among the most studied signaling proteins and represent promising therapeutic targets for human neoplastic disease. Despite the high level of interest in these proteins, direct analysis of most aspects of Ras protein biology in living cells has not been possible, because m...

journal_title：Methods in enzymology

authors： Westwick JK,Michnick SW

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：:IF-liposomes are formed by a unique process that involves fusing small liposomes into interdigitated lipid sheets, using either ethanol or hydrostatic pressure. The interdigitation-fusion method requires liposome formulations with lipids that form the L beta I phase. Preparing ethanol-induced IF-liposomes is simple an...

journal_title：Methods in enzymology

authors： Ahl PL,Perkins WR

更新日期：2003-01-01 00:00:00

abstract：:Isothermal titration calorimetry (ITC) provides a sensitive and accurate means by which to study the thermodynamics of binding reactions. In addition, it enables label-free measurement of enzymatic reactions. The advent of extremely sensitive microcalorimeters have made it increasingly valuable as a tool for hit valid...

journal_title：Methods in enzymology

authors： Recht MI,Nienaber V,Torres FE

更新日期：2016-01-01 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

authors： O'Prey J,Sakamaki J,Baudot AD,New M,Van Acker T,Tooze SA,Long JS,Ryan KM

更新日期：2017-01-01 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：:Peroxiredoxins (Prdxs) are a family of proteins which catalyze the reduction of H2O2 through the interaction of active site cysteine residues. Conserved within all plant and animal kingdoms, the function of these proteins is related to protection from oxidation or participation of signaling through degradation of H2O2...

journal_title：Methods in enzymology

authors： Ambruso DR

更新日期：2013-01-01 00:00:00

abstract：:Cell proliferation requires the coordination of multiple signaling pathways as well as the provision of metabolic substrates. Nutrients are required to generate such building blocks and their form of utilization differs to significant extents between malignant tissues and their nontransformed counterparts. Thus, oncog...

journal_title：Methods in enzymology

authors： Iurlaro R,León-Annicchiarico CL,Muñoz-Pinedo C

更新日期：2014-01-01 00:00:00