Leucine-tRNA ligase complexes.

abstract：:In the yeast Saccharomyces cerevisiae, autophagy contributes to the sustaining of cell viability under starvation conditions, possibly through the supply of amino acids that is generated as a result of the degradation of cytosolic materials. Therefore, cellular viability is one of the best indexes for monitoring the c...

journal_title：Methods in enzymology

authors： Noda T

更新日期：2008-01-01 00:00:00

abstract：:Osmoregulation is the active control of the cellular water balance and encompasses homeostatic mechanisms crucial for life. The osmoregulatory system in the yeast Saccharomyces cerevisiae is particularly well understood. Key to yeast osmoregulation is the production and accumulation of the compatible solute glycerol, ...

journal_title：Methods in enzymology

authors： Hohmann S,Krantz M,Nordlander B

更新日期：2007-01-01 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：:Nonribosomal peptide biosynthesis is a complex enzymatic assembly responsible for producing a great diversity of bioactive peptide natural products. Due to the recurring arrangement of catalytic domains within these machineries, great interest has been shown in reengineering these pathways to produce novel, designer p...

journal_title：Methods in enzymology

authors： Greule A,Charkoudian LK,Cryle MJ

更新日期：2019-01-01 00:00:00

abstract：:Malaria represents an emerging disease because of increasing parasite resistance against available drugs and because of increasing geographical distribution of the causative agent, Plasmodium falciparum. The complete genome of Plasmodium was sequenced recently, revealing that the parasite harbors only one glutathione ...

journal_title：Methods in enzymology

authors： Deponte M,Becker K

更新日期：2005-01-01 00:00:00

abstract：:Primary mixed liver cells were isolated from rats that had been fed an amino acid (AA) diet in which natural protein was replaced with a defined mixture of pure AAs. Nitric oxide (NO) production from these cells in vitro was monitored using a nitric oxide (NO)-selective fluorescent probe, diaminofluorescein, followed ...

journal_title：Methods in enzymology

authors： Kotake Y,Kishida H,Nakae D,Floyd RA

更新日期：2005-01-01 00:00:00

abstract：:After cell lysis, the most often used second step in a protein purification procedure is some sort of a rapid, bulk precipitation step. This is commonly accomplished by altering the solvent conditions and taking advantage of the changes in solubility of your protein of interest relative to those of many of the other p...

journal_title：Methods in enzymology

authors： Burgess RR

更新日期：2009-01-01 00:00:00

abstract：:Gas chromatography-mass spectrometry (GC-MS) is one of the three most popular analytical platforms for metabolomics and is largely employed for the study of oncometabolism. Large volumes of data are usually generated in a GC-MS experiment, and many analytical steps are required to extract biologically relevant informa...

journal_title：Methods in enzymology

authors： Koo I,Wei X,Zhang X

更新日期：2014-01-01 00:00:00

abstract：:The methods of computational chemistry and molecular modeling are becoming more and more accessible to biochemists with the advent of fast, inexpensive graphics workstations and well-tested computer programs. The state of the art in small molecules allows chemists to use these programs as "black boxes" and be confiden...

journal_title：Methods in enzymology

authors： Wampler JE

更新日期：1994-01-01 00:00:00

abstract：:A central goal of biology is to understand how transcription factors target and regulate specific genes and networks to control cell fate and function. An equally important goal of synthetic biology, chemical biology, and personalized medicine is to devise molecules that can regulate genes and networks in a programmab...

journal_title：Methods in enzymology

authors： Tietjen JR,Donato LJ,Bhimisaria D,Ansari AZ

更新日期：2011-01-01 00:00:00

abstract：:G-protein-coupled receptors (GPCRs) represent a major class of receptors through which a number of signals ranging from photons to large glycoprotein hormones are recognized. Human genome encodes about 800 GPCRs, yet very little structural information is available on this class of receptors. Structural studies provide...

journal_title：Methods in enzymology

authors： Ashok Y,Nanekar RT,Jaakola VP

更新日期：2013-01-01 00:00:00

abstract：:Omics approaches have become popular in biology as powerful discovery tools, and currently gain in interest for diagnostic applications. Establishing the accurate genome sequence of any organism is easy, but the outcome of its annotation by means of automatic pipelines remains imprecise. Some protein-encoding genes ma...

journal_title：Methods in enzymology

authors： Armengaud J

更新日期：2017-01-01 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 Escherichia coli fumarate and nitrate reductase (FNR) regulator protein is an important transcriptional regulator that controls the expression of a large regulon of more than 100 genes in response to changes in oxygen availability. FNR is active when it acquires a [4Fe-4S](2+) cluster under anaerobic conditions. T...

journal_title：Methods in enzymology

authors： Crack JC,Le Brun NE,Thomson AJ,Green J,Jervis AJ

更新日期：2008-01-01 00:00:00

abstract：:Mass spectrometry (MS) has gradually replaced classical methods as a major tool in protein sequencing and characterization. However, the sample preparation repertoire has not changed very much; it has just been adjusted to the needs of the new analytical method. In this chapter frequently used in-solution enzymatic di...

journal_title：Methods in enzymology

authors： Medzihradszky KF

更新日期：2005-01-01 00:00:00

abstract：:Life cell imaging is a tool for cell biology that has provided invaluable insights into many dynamic processes such as cell division, morphogenesis, or endo- and exocytosis. While observing cells by time-lapse imaging is a standard procedure in many systems, this technique was until recently not available for blood st...

journal_title：Methods in enzymology

authors： Grüring C,Spielmann T

更新日期：2012-01-01 00:00:00

abstract：:Genetically encoded fluorescent sensors are essential tools in modern biological research, and recent advances in fluorescent proteins (FPs) have expanded the scope of sensor design and implementation. In this review we compare different sensor platforms, including Förster resonance energy transfer (FRET) sensors, flu...

journal_title：Methods in enzymology

authors： Sanford L,Palmer A

更新日期：2017-01-01 00:00:00

abstract：:This protocol describes a method for determining the concentration of a nucleic acid sample. ...

journal_title：Methods in enzymology

authors： Kolitz S,Lorsch J

更新日期：2013-01-01 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 Ras gene family has been implicated in the development of many human epithelial cancers. Mutations in K-ras or its downstream mediator BRAF have been detected in about two thirds of low-grade serous carcinomas and borderline serous tumors; mutations in K-ras are also often present in benign and invasive mucinous o...

journal_title：Methods in enzymology

authors： Rosen DG,Yang G,Bast RC Jr,Liu J

更新日期：2006-01-01 00:00:00

abstract：:Proteins change their charge state through protonation and redox reactions as well as through binding charged ligands. The free energy of these reactions is dominated by solvation and electrostatic energies and modulated by protein conformational relaxation in response to the ionization state changes. Although computa...

journal_title：Methods in enzymology

authors： Gunner MR,Baker NA

更新日期：2016-01-01 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：: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

authors： Locker N,Lukavsky PJ

更新日期：2007-01-01 00:00:00

abstract：:A neuron is able to seamlessly respond to a number of signals, in a timely and specific manner. This process, of integrating multiple inputs, relays on the orchestration of intracellular events by signaling networks. The inherent complexity of signaling networks has made computational modeling a useful approach to und...

journal_title：Methods in enzymology

authors： Wenderski WC,Neves SR

更新日期：2012-01-01 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：:Natural products continue to demonstrate their utility both as therapeutics and as molecular probes for the discovery and mechanistic deconvolution of various cellular processes. However, this utility is dampened by the inherent difficulties involved in isolating and characterizing new bioactive natural products, in o...

journal_title：Methods in enzymology

authors： Low WK,Dang Y,Schneider-Poetsch T,Shi Z,Choi NS,Rzasa RM,Shea HA,Li S,Park K,Ma G,Romo D,Liu JO

更新日期：2007-01-01 00:00:00

abstract：:Adiponectin is a circulating bioactive hormone secreted by adipocytes as oligomers ranging in size from 90 kDa trimers and 180 kDa hexamers to larger high molecular weight oligomers that may reach 18- or 36-mers in size. While total circulating adiponectin levels correlate well with metabolic health, it is the relativ...

journal_title：Methods in enzymology

authors： Rutkowski JM,Scherer PE

更新日期：2014-01-01 00:00:00

abstract：:Experimental screening for protein-ligand interactions is a central task in drug discovery. Nuclear magnetic resonance (NMR) spectroscopy enables the determination of binding affinities, as well as the measurement of structural and dynamic parameters governing the interaction. With traditional liquid-state NMR relying...

journal_title：Methods in enzymology

authors： Kim Y,Hilty C

更新日期：2019-01-01 00:00:00

abstract：:The goal of the signal transduction pathways, such as those controlled by Ras, is in large part to ensure highly stringent regulation of the target genes in the nucleus, which are collectively responsible for the signal output, or phenotypes, of the cell. Understanding of the Ras effect ultimately requires the identif...

journal_title：Methods in enzymology

authors： Jo H,Cho YJ,Zhang H,Liang P

更新日期：2001-01-01 00:00:00

abstract：:Retinoic acid (RA), the biologically active metabolite of vitamin A, regulates a vast spectrum of biological processes, such as cell differentiation, proliferation, apoptosis, and morphogenesis. microRNAs (miRNAs) play a crucial role in regulating gene expression by binding to messenger RNA (mRNA) which leads to mRNA ...

journal_title：Methods in enzymology

authors： Wang L,Rohatgi AP,Wan YY

更新日期：2020-01-01 00:00:00