S-glutathionylation of glyceraldehyde-3-phosphate dehydrogenase: role of thiol oxidation and catalysis by glutaredoxin.

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：:Immunogenic cell death (ICD), induced by certain anticancer chemotherapeutics, leads to the emission of danger associated molecular patterns (DAMP) by cancer cells, which facilitates the attraction, activation and maturation of dendritic cells (DC) as well as the subsequent priming of effector T cells. In this context...

journal_title：Methods in enzymology

authors： Cerrato G,Liu P,Martins I,Kepp O,Kroemer G

更新日期：2020-01-01 00:00:00

abstract：:Classifications and characterizations of specific proteins, such as enzymes, not only allow us to understand biosynthetic and metabolic pathways but they also help to drive our understanding of protein structure and function. How those characterizations are evaluated, however, may change our interpretations and lead u...

journal_title：Methods in enzymology

authors： Bradshaw HB,Leishman E

更新日期：2017-01-01 00:00:00

abstract：:Monte Carlo simulations of neighbor exclusion models have been used to demonstrate the importance of collecting and fitting data over a wide range of saturation. Low saturation data are important for good estimates of the affinity K of a drug or protein for the lattice site. High saturation data are important for dist...

journal_title：Methods in enzymology

authors： Correia JJ,Chaires JB

更新日期：1994-01-01 00:00:00

abstract：:It is widely acknowledged that mitochondria are highly active structures that rapidly respond to cellular and environmental perturbations by changing their shape, number, and distribution. Mitochondrial remodeling is a key component of diverse biological processes, ranging from cell cycle progression to autophagy. In ...

journal_title：Methods in enzymology

authors： Marchi S,Bonora M,Patergnani S,Giorgi C,Pinton P

更新日期：2017-01-01 00:00:00

abstract：:We have developed a genetic system, termed IVET (in vivo expression technology), designed to identify bacterial genes that are induced when a pathogen infects its host. A subset of these induced genes should include those that encode virulence factors, products specifically required for the infection process. The syst...

journal_title：Methods in enzymology

authors： Slauch JM,Mahan MJ,Mekalanos JJ

更新日期：1994-01-01 00:00:00

abstract：:The three major considerations in reducing metal-gel binding are (1) the type of gel to be used, (2) the ionic strength of the eluent, and (3) alkaline reduction of the gel. Each of these parameters should be considered in advance of an experiment. A method is offered for determining the optimal gel type, gel treatmen...

journal_title：Methods in enzymology

authors： Martin MT

更新日期：1988-01-01 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：:There is a great need for practicing scientists to volunteer their time and expertise in the K-12th grade science classroom. We have found that bioluminescence is a fun and exciting way to teach basic science concepts and is an excellent tool for the volunteering scientist. We have had very positive reactions from bot...

journal_title：Methods in enzymology

authors： Hammer M,Andrade JD

更新日期：2000-01-01 00:00:00

abstract：:Virtually all eukaryotic cell functions and signaling pathways are regulated by protein phosphorylation. The Rad53 kinase plays crucial roles in the DNA damage response in Saccharomyces cerevisiae and is widely used as a surrogate marker for DNA damage checkpoint activation by diverse genotoxic agents. Most currently ...

journal_title：Methods in enzymology

authors： Hoch NC,Chen ES,Tsai MD,Heierhorst J

更新日期：2017-01-01 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：: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

abstract：:Isothermal Titration Calorimetry (ITC) provides a sensitive and accurate means by which to study the thermodynamics of RNA folding, RNA binding to small molecules, and RNA-protein interactions. The advent of extremely sensitive instrumentation and the increasing availability of ITC in shared facilities have made it in...

journal_title：Methods in enzymology

authors： Feig AL

更新日期：2009-01-01 00:00:00

abstract：:Use of fluorescence in biology and biochemistry for imaging and characterizing equilibrium and dynamic processes is growing exponentially. Much progress has been made in the last few years on the microscopic understanding of the underlying principles of what controls the wavelength and quenching of fluorescence in bio...

journal_title：Methods in enzymology

authors： Callis PR

更新日期：2011-01-01 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：:Although most eukaryotic mRNAs are degraded by exonucleases acting on either end of the molecule, a subset of mRNAs undergo endonuclease cleavage within the mRNA body. Endonuclease cleavage can be activated by cellular stress, extracellular signals, or by ribosome stalling, as might occur at a premature termination co...

journal_title：Methods in enzymology

authors： Otsuka Y,Schoenberg DR

更新日期：2008-01-01 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：:Antibodies will be immobilized on a cyanogen bromide-activated Sepharose for subsequent use in pull-down assays or immunoaffinity purification. ...

journal_title：Methods in enzymology

authors： Kavran JM,Leahy DJ

更新日期：2014-01-01 00:00:00

abstract：:Protein-protein interactions (PPIs) are universal to life and their study and understanding is critical to drug discovery and bioengineering efforts. Historically, X-ray crystallography, isothermal titration calorimetry and other biophysical methods have been used to study PPIs, but can be costly and are low throughpu...

journal_title：Methods in enzymology

authors： Charov K,Burkart MD

更新日期：2020-01-01 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：:A variety of recent advances in single-molecule methods are now making possible the routine measurement of the distinct catalytic trajectories of individual enzymes. Unlike their bulk counterparts, these measurements directly reveal the fluctuations inherent to enzymatic dynamics, and statistical measures of these flu...

journal_title：Methods in enzymology

authors： Moffitt JR,Chemla YR,Bustamante C

更新日期：2010-01-01 00:00:00

abstract：:Single-molecule Förster resonance energy transfer (smFRET) is a versatile tool for studying biomolecules in a quantitative manner. Multiple conformations within and interactions between biomolecules can be detected and their kinetics can be determined. Thus, smFRET has become an essential tool in enzymology. Ordinary ...

journal_title：Methods in enzymology

authors： Götz M,Wortmann P,Schmid S,Hugel T

更新日期：2016-01-01 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：: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：:ADP-ribosylation is a protein post-translational modification that is critically involved in a wide array of biological processes connected to cell stress responses. Enzymes known as poly-ADP-ribose polymerases (PARPs) catalyze the addition of the ADP-ribose units to amino acids with various side chain chemistries. In...

journal_title：Methods in enzymology

authors： Li P,Zhen Y,Yu Y

更新日期：2019-01-01 00:00:00

abstract：:Protein oxidation is implicated in atherogenesis and other inflammatory conditions. Measuring levels of chlorinated and nitrated proteins in biological matrices serves as a quantitative index of oxidative stress in vivo. One potential mechanism for oxidative stress involves myeloperoxidase, a heme protein expressed by...

journal_title：Methods in enzymology

authors： Shao B,Heinecke JW

更新日期：2008-01-01 00:00:00

abstract：:G-protein-coupled receptors (GPCRs) initiate signaling pathways via trimetric guanine nucleotide-binding proteins. GPCRs are classified based on their ligand-binding properties and molecular phylogenetic analyses. Nonetheless, these later analyses are in most case dependent on multiple sequence alignments, themselves ...

journal_title：Methods in enzymology

authors： Rehman ZU,Mirza MT,Khan A,Xhaard H

更新日期：2013-01-01 00:00:00

abstract：:Posttranslational modification of circadian clock proteins by phosphorylation is an essential regulatory process in the control of eukaryotic circadian clocks. In the Neurospora circadian clock, the key clock protein FREQUENCY (FRQ) is progressively phosphorylated. The phosphorylation of FRQ is regulated by both kinas...

journal_title：Methods in enzymology

authors： Liu Y

更新日期：2005-01-01 00:00:00

abstract：:We describe here the methods we have used to generate selective peptide inhibitors and activators of PKC-mediated signaling. These approaches should be applicable to any signaling event that is dependent on protein-protein interaction. Furthermore, targeting downstream enzymes in signal transduction has been notorious...

journal_title：Methods in enzymology

authors： Schechtman D,Mochly-Rosen D

更新日期：2002-01-01 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