Biochemical Reconstitution and Spectroscopic Analysis of Iron-Sulfur Proteins.

abstract：:In recent years the facile, yet powerful, genetics of the baker's yeast Saccharomyces cerevisiae has been appropriated for the study of amyloid toxicity. Several models of amyloid toxicity using this simple eukaryotic organism have been developed that faithfully recapitulate many disease-relevant phenotypes. Furthermo...

journal_title：Methods in enzymology

authors： Giorgini F,Muchowski PJ

更新日期：2006-01-01 00:00:00

abstract：:Since the onset of the genomic era more than 1000 bacterial genomes have been sequenced and several fold more are expected to be completed in the near future. These genome sequences supply a wealth of information that can be exploited by statistical methods to gain significant insights into cellular processes. In Volu...

journal_title：Methods in enzymology

authors： Lunt B,Szurmant H,Procaccini A,Hoch JA,Hwa T,Weigt M

更新日期：2010-01-01 00:00:00

abstract：:The group IVA phospholipase A2, commonly called cytosolic phospholipase A2α (cPLA2α), is a widely expressed enzyme that hydrolyzes membrane phospholipid to produce arachidonic acid and lysophospholipids, which are precursors for a number of bioactive lipid mediators. Arachidonic acid is metabolized through the cycloox...

journal_title：Methods in enzymology

authors： Yun B,Leslie CC

更新日期：2017-01-01 00:00:00

abstract：:The neutral arachidonic acid derivatives N-arachidonoyl ethanolamine (anandamide or AEA), and 2-arachidonoylglycerol (2-AG) have been identified as endogenous ligands for the cannabinoid receptors. Additionally, these compounds have been identified as substrates of the second isoform of the cyclooxygenase enzyme (COX-...

journal_title：Methods in enzymology

authors： Kingsley PJ,Marnett LJ

更新日期：2007-01-01 00:00:00

abstract：:Inducible expression systems represent the founding technology for the emergence of synthetic biology in mammalian cells. The core molecules in these systems are bacterial regulator proteins that bind to or dissociate from a cognate DNA operator sequence in response to an exogenous stimulus like a small-molecule induc...

journal_title：Methods in enzymology

authors： Karlsson M,Weber W,Fussenegger M

更新日期：2011-01-01 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：:Genomes can be viewed in terms of their gene content and the order in which the genes appear along each chromosome. Evolutionary events that affect the gene order or content are "rare genomic events" (rarer than events that affect the composition of the nucleotide sequences) and have been advocated by systematists for...

journal_title：Methods in enzymology

authors： Moret BM,Warnow T

更新日期：2005-01-01 00:00:00

abstract：:Autophagy is an ancient cell survival pathway that is induced by metabolic stress and that helps prevent bioenergetic failure. This pathway has emerged as a promising new target in cancer treatment, where agents that inhibit autophagic degradation have efficacy in preventing cancer and in treating resistant disease wh...

journal_title：Methods in enzymology

authors： Dorsey FC,Steeves MA,Prater SM,Schröter T,Cleveland JL

更新日期：2009-01-01 00:00:00

abstract：:During the past several years, major progress has been accomplished in the production of "designer cellulosomes," artificial enzymatic complexes that were demonstrated to efficiently degrade crystalline cellulose. This progress is part of a global attempt to promote biomass waste solutions and biofuel production. In d...

journal_title：Methods in enzymology

authors： Vazana Y,Moraïs S,Barak Y,Lamed R,Bayer EA

更新日期：2012-01-01 00:00:00

abstract：:Ubiquitin-like protein (Ubl) ligation is common to diverse archaea and targets many cellular pathways, including those associated with sulfur mobilization, and also tags proteins as substrates for degradation by the proteasome. Here we highlight protocols to assay proteasome function and Ubl ligation in archaea. A cha...

journal_title：Methods in enzymology

authors： Fu X,Adams Z,Maupin-Furlow J

更新日期：2019-01-01 00:00:00

abstract：:The monitoring of trace amounts of nitric oxide (NO) is of great interest for biomedical applications. High-resolution infrared spectroscopy, like laser magnetic resonance spectroscopy (LMRS), is a versatile technique for the quantitative and isotope-selective analysis of low concentrations of NO. The ability to disti...

journal_title：Methods in enzymology

authors： Gäbler R,Lehmann J

更新日期：2005-01-01 00:00:00

abstract：:This chapter presents methodologies for RNA extraction from soils coupled with competitive reverse transcription-polymerase chain reaction and capillary electrophoresis techniques. Combined, these approaches provide new capabilities to quantify gene expression in different environments and can aid our understanding of...

journal_title：Methods in enzymology

authors： Semrau JD,Han JI

更新日期：2005-01-01 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

authors： Winkler DD,Luger K,Hieb AR

更新日期：2012-01-01 00:00:00

abstract：:The diverse structures of polyketide natural products are reflected by the equally diverse polyketide biosynthetic enzymes, namely polyketide synthases (PKSs). Three major classes of PKSs are known-noniterative type I PKSs, iterative type II PKSs and acyl carrier protein-independent type III PKSs, each of which consis...

journal_title：Methods in enzymology

authors： Cheng YQ,Coughlin JM,Lim SK,Shen B

更新日期：2009-01-01 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：:Two factors, climate change brought on by rising atmospheric CO2 levels and the accelerating shift toward renewable energy sources, have together worked to heighten interest in understanding how biological catalysts so effectively bring about the reduction of CO2 to formate, with potential applications for both biorem...

journal_title：Methods in enzymology

authors： Niks D,Hille R

更新日期：2018-01-01 00:00:00

abstract：:Human embryonic stem cells (hESCs) are derived from preimplantation embryos. Approximately 60% of human embryos are blocked during in vitro development. Although statistics are inconclusive, experience demonstrates that hESCs are more effectively derived from high-quality embryos. In this way, optimal human embryo cul...

journal_title：Methods in enzymology

authors： Mercader A,Valbuena D,Simón C

更新日期：2006-01-01 00:00:00

abstract：:The Omega class of cytosolic glutathione transferases was initially recognized by bioinformatic analysis of human sequence databases, and orthologous sequences were subsequently discovered in mouse, rat, pig, Caenorhabditis elegans, Schistosoma mansoni, and Drosophila melanogaster. In humans and mice, two GSTO genes h...

journal_title：Methods in enzymology

authors： Whitbread AK,Masoumi A,Tetlow N,Schmuck E,Coggan M,Board PG

更新日期：2005-01-01 00:00:00

abstract：:A fundamental concept of phosphorylation-mediated signaling is the precise switching between discrete functional conformations. According to the traditional view, phosphorylation induces a new, active conformation. In this chapter, a series of NMR experiments performed on a response regulator are described that challe...

journal_title：Methods in enzymology

authors： Gardino AK,Kern D

更新日期：2007-01-01 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：: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：:Extracellular thiol/disulfide redox environments are highly regulated in healthy individuals and become oxidized in disease. This oxidation affects the function of cell surface receptors, ion channels, and structural proteins. Downstream signaling due to changes in extracellular redox potential can be studied using a ...

journal_title：Methods in enzymology

authors： Go YM,Jones DP

更新日期：2010-01-01 00:00:00

abstract：:In the least-squares fitting of data, there is a unique answer to the question of how the data should be weighted: inversely as their variance. Any other weighting gives less than optimal efficiency and leads to unreliable estimates of the parameter uncertainties. In calibration, knowledge of the data variance permits...

journal_title：Methods in enzymology

authors： Tellinghuisen J

更新日期：2009-01-01 00:00:00

abstract：:Powerful specialized software is essential for managing, quantifying, and ultimately deriving scientific insight from results of a microarray experiment. We have developed a suite of software applications, known as TM4, to support such gene expression studies. The suite consists of open-source tools for data managemen...

journal_title：Methods in enzymology

authors： Saeed AI,Bhagabati NK,Braisted JC,Liang W,Sharov V,Howe EA,Li J,Thiagarajan M,White JA,Quackenbush J

更新日期：2006-01-01 00:00:00

abstract：:There is currently no known genetic disease linked to prolactin (PRL) or its receptor (PRLR) in humans. Recently, we identified three missense variants of the PRLR in patients presenting with breast tumors. Two of them (named PRLR(I146L) and PRLR(I76V)) had been reported earlier, but failed to draw much attention beca...

journal_title：Methods in enzymology

authors： Goffin V,Bogorad RL,Touraine P

更新日期：2010-01-01 00:00:00

abstract：:Intramembrane-cleaving proteases (I-CLiPs) are a group of membrane-associated proteases with a unique feature: they are believed to cleave their substrate within the hydrophobic lipid bilayer, even though peptide bond hydrolysis requires a water molecule. Escherichia coli RseP, which belongs to the S2P zinc metallopro...

journal_title：Methods in enzymology

authors： Hizukuri Y,Akiyama K,Akiyama Y

更新日期：2017-01-01 00:00:00

abstract：:When isolating ribonucleoprotein (RNP) complexes by an affinity selection approach, tagging the RNA component can prove to be strategically important. This is especially true for purifying single types of snoRNPs, because in most cases the snoRNA is thought to be the only unique component. Here, we present a general s...

journal_title：Methods in enzymology

authors： Piekna-Przybylska D,Liu B,Fournier MJ

更新日期：2007-01-01 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

authors： Kerber T,Vrielink A

更新日期：2020-01-01 00:00:00

abstract：:The human cannabinoid receptor, CB1, has been difficult to purify in a functional form, hampering structural and biophysical studies. Here, we present our approaches for obtaining pure, detergent solubilized, functional CB1. We also discuss our site-directed fluorescence labeling (SDFL) methods for identifying differe...

journal_title：Methods in enzymology

authors： Fay JF,Farrens DL

更新日期：2017-01-01 00:00:00

abstract：:Complex RNA structures regulate many biological processes but are often too large for structure determination by nuclear magnetic resonance (NMR) methods. We determined the solution structure of domain II of the hepatitis C viral internal ribosome entry site (HCV IRES), a 25-kDa RNA, using a novel NMR approach. Conven...

journal_title：Methods in enzymology

authors： Lukavsky PJ,Puglisi JD

更新日期：2005-01-01 00:00:00