A Networks Approach to Modeling Enzymatic Reactions.

abstract：:Changes in chromatin structure emanating from DNA breaks are among the most initiating events in the damage response of the cell. In higher eukaryotes, poly(ADP-ribose) polymerase-1 (PARP-1) translates the occurrence of DNA breaks detected by its zinc-finger domain into a signal, poly ADP-ribose, synthesized and ampli...

journal_title：Methods in enzymology

authors： Dantzer F,Amé JC,Schreiber V,Nakamura J,Ménissier-de Murcia J,de Murcia G

更新日期：2006-01-01 00:00:00

abstract：:Chloroplast genomes of land plants and algae contain generally between 100 and 150 genes. These genes are involved in plastid gene expression and photosynthesis and in various other tasks. The function of some chloroplast genes is still unknown and some of them appear to be essential for growth and survival. Repressib...

journal_title：Methods in enzymology

authors： Ramundo S,Rochaix JD

更新日期：2015-01-01 00:00:00

abstract：:Although the use of monoclonal antibodies to characterize the Ia antigen system is still in its infancy, the usefulness of this approach is clearly apparent. Since xenogeneic immunization can be used to generate these antibodies, the chances are good that a greater variety of Ia types and determinants will be detected...

journal_title：Methods in enzymology

authors： Zweig SE,Shevach EM

更新日期：1983-01-01 00:00:00

abstract：:Oxidized phospholipids play an important role in execution of the mitochondrial stage of apoptosis and clearance of apoptotic cells by macrophages. Therefore, the identification and quantification of oxidized phospholipids generated during apoptosis are very important. These can be achieved successfully by a newly dev...

journal_title：Methods in enzymology

authors： Tyurin VA,Tyurina YY,Kochanek PM,Hamilton R,DeKosky ST,Greenberger JS,Bayir H,Kagan VE

更新日期：2008-01-01 00:00:00

abstract：:Scanning transmission electron microscopy (STEM) of unstained, freeze-dried biological macromolecules in the dark-field mode provides an image based on the number of electrons elastically scattered by the constituent atoms of the macromolecule. The image of each isolated particle provides information about the project...

journal_title：Methods in enzymology

authors： Wall JS,Simon MN,Lin BY,Vinogradov SN

更新日期：2008-01-01 00:00:00

abstract：:This chapter reviews basic concepts of nonlinear fluorescence upconversion, a technique whose temporal resolution is essentially limited only by the pulse width of the ultrafast laser. Design aspects for upconversion spectrophotofluorometers are discussed, and a recently developed system is described. We discuss appli...

journal_title：Methods in enzymology

authors： Xu J,Knutson JR

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 remarkable success of cancer immunotherapies, especially the checkpoint blocking antibodies, in a subset of patients has reinvigorated the study of tumor-immune crosstalk and its role in heterogeneity of response. High-throughput sequencing and imaging technologies can help recapitulate various aspects of the tumo...

journal_title：Methods in enzymology

authors： Bhinder B,Elemento O

更新日期：2020-01-01 00:00:00

abstract：:Proteins are fragile molecules that often require great care during purification to ensure that they remain intact and fully active. Nowadays, many proteins are also purified in small amounts under denaturing conditions by various gel electrophoretic techniques, such that inactive proteins are obtained. But even here,...

journal_title：Methods in enzymology

authors： Deutscher MP

更新日期：2009-01-01 00:00:00

abstract：:We describe here the components and uses of two image intensifier systems. The SIT camera system is convenient, relatively inexpensive, readily adaptable to most microscopes, and reliable. It suffers from lack of resolution at high gain levels and the inability to extend its sensitivity by accumulating an image over t...

journal_title：Methods in enzymology

authors： Willingham MC,Pastan IH

更新日期：1983-01-01 00:00:00

abstract：:One of the most widely employed methods in metagenomics is the amplification and sequencing of the highly conserved ribosomal RNA (rRNA) genes from organisms in complex microbial communities. rRNA surveys, typically using the 16S rRNA gene for prokaryotic identification, provide information about the total diversity a...

journal_title：Methods in enzymology

authors： Preheim SP,Perrotta AR,Friedman J,Smilie C,Brito I,Smith MB,Alm E

更新日期：2013-01-01 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：:Differential scanning calorimetry (DSC) detects protein thermal unfolding by directly measuring the heat absorbed. Simple DSC experiments that require relatively small amounts of pure material can provide a wealth of information related to structure, especially with respect to domain architecture, without the need for...

journal_title：Methods in enzymology

authors： Yang Z,Brouillette CG

更新日期：2016-01-01 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：: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：:Ultraviolet photodissociation time-of-flight (TOF) mass spectrometry (MS) is described as a method for determination of peptide ion primary structure. Monoisotopic selection and bond-specific activation, combined with the rapidity of TOF MS analysis, render this technique invaluable to the rapidly expanding field of p...

journal_title：Methods in enzymology

authors： Morgan JW,Hettick JM,Russell DH

更新日期：2005-01-01 00:00:00

abstract：:PRA2 was found to interact with the ER-localized protein VAMP-associated protein of 33 kDa or VAP-33 by a yeast two-hybrid screen. We describe here the purification of PRA2 and VAP-33 as well as an in vitro pull-down procedure to verify the interaction. PRA2 was found to form a large sodium dodecyl sulfate (SDS)-insol...

journal_title：Methods in enzymology

authors： Gougeon PY,Ngsee JK

更新日期：2005-01-01 00:00:00

abstract：:A procedure for an in vitro signaling assay is described for the MAPK and NFkappaB pathways. The method uses a membrane-cleared lysate that contains all the soluble components required for activating these signaling cascades. The pathways can be activated by variety of molecules, including kinases, G-proteins, and E3 ...

journal_title：Methods in enzymology

authors： Mukherjee S,Orth K

更新日期：2008-01-01 00:00:00

abstract：:Genetic modifier screens offer a powerful, indeed a uniquely powerful tool for the analysis and identification of elements capable of modulating specific cellular functions in development. Here, we describe the methodology that allowed us to explore the genetic circuitry that affects a Notch mutant phenotype caused by...

journal_title：Methods in enzymology

authors： Hori K,Sen A,Artavanis-Tsakonas S

更新日期：2014-01-01 00:00:00

abstract：:RNA modifications, with over one hundred known so far, are commonly proposed to fine-tune the structure and function of RNA. While modifications in rRNA and tRNA are used to modulate RNA folding and decoding properties, little is known about the function of internal modifications in mRNA/lncRNA, which includes N(6)-me...

journal_title：Methods in enzymology

authors： Liu N,Pan T

更新日期：2015-01-01 00:00:00

abstract：:Enzyme immobilization is a widespread empiric technology to achieve more stable, active and reusable enzymes. The empiricism can be reduced by the application of rational design procedures employing bioinformatic tools, engineered-proteins and detailed analyses of existent data. In this chapter, we describe relevant a...

journal_title：Methods in enzymology

authors： Sastre DE,Reis EA,Marques Netto CGC

更新日期：2020-01-01 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：:The characterization of microbial adhesins is greatly facilitated by the use of synthetic peptides. Synthetic peptides can be used to identify specific antigenic epitopes, to delineate receptor-binding domain of adhesins, and to facilitate the characterization of the adhesin, and they allow for a direct examination of...

journal_title：Methods in enzymology

authors： Lee KK,Wong WY,Sheth HB,Hodges RS,Paranchych W,Irvin RT

更新日期：1995-01-01 00:00:00

abstract：:Chaperone-mediated autophagy (CMA) is a selective type of autophagy responsible for the lysosomal degradation of soluble cytosolic proteins. In contrast to other forms of autophagy where cargo is sequestered and delivered to lysosomes through membrane fusion/excision, CMA substrates reach the lysosomal lumen after dir...

journal_title：Methods in enzymology

authors： Kaushik S,Cuervo AM

更新日期：2009-01-01 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：:Acidic organelles and vesicles, such as endosomes, lysosomes, autophagosomes, trans-Golgi network, and synaptic vesicles, are known to play important roles in a broad range of cellular events. To facilitate studying these multifunctional systems, we describe here an acid-brightening fluorescent protein (abFP), which f...

journal_title：Methods in enzymology

authors： Wang N,Wang L

更新日期：2020-01-01 00:00:00

abstract：:Hydrogen peroxide (H2O2) is a ubiquitous biological molecule whose wide range of biological functions depends on its concentration. In this chapter, we compare the delivery of H2O2 to cells as (1) a single initial dose (bolus addition); (2) a continuous source using, for example, glucose oxidase; and (3) a steady stat...

journal_title：Methods in enzymology

authors： Marinho HS,Cyrne L,Cadenas E,Antunes F

更新日期：2013-01-01 00:00:00

abstract：:Lysosomes are specialized organelles for protein recycling and as such are involved in the terminal steps of autophagy. However, it has become evident that lysosomes also play an important role in the progression of apoptosis. This latter function seems to be dependent on lysosomal proteases, which need to be released...

journal_title：Methods in enzymology

authors： Ivanova S,Repnik U,Bojic L,Petelin A,Turk V,Turk B

更新日期：2008-01-01 00:00:00

abstract：:Circadian clocks are present in most cells and are essential for maintenance of daily rhythms in physiology, mood, and cognition. Thus, not only neurons of the central circadian pacemaker but also many other peripheral tissues possess the same functional and self-sustained circadian clocks. Surprisingly, however, thei...

journal_title：Methods in enzymology

authors： Gaspar L,Brown SA

更新日期：2015-01-01 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