Regulation of cancer metabolism by oncogenes and tumor suppressors.

abstract：:Hypoxia and inflammation are coincidental events in an array of diseased tissues, including chronically inflamed sites (e.g., inflammatory bowel disease, rheumatoid arthritis), growing tumors, myocardial infarcts, atherosclerotic plaques, healing wounds, and sites of bacterial infection (Murdoch et al., 2005). An unde...

journal_title：Methods in enzymology

authors： Cummins EP,Comerford KM,Scholz C,Bruning U,Taylor CT

更新日期：2007-01-01 00:00:00

abstract：:In this chapter we have detailed a method that can be generalized to link virtually any DNA substrate to a chromatography matrix at its ends via an avidin-biotin linkage. We have used this technique to construct a left-handed Z-DNA column for the purpose of identification and purification of Z-DNA-binding proteins. Th...

journal_title：Methods in enzymology

authors： Fishel R,Anziano P,Rich A

更新日期：1990-01-01 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：:Bile acids are converted to their glycine and taurine N-acyl amidates by enzymes in the liver in a two-step process. This conjugation reaction increases the aqueous solubility of bile acids, particularly in the acidic environment of the initial portion of the small intestine. In the first step, bile acid coenzyme A (C...

journal_title：Methods in enzymology

authors： Shonsey EM,Wheeler J,Johnson M,He D,Falany CN,Falany J,Barnes S

更新日期：2005-01-01 00:00:00

abstract：:In this chapter, we describe a set of complementary techniques that we use to study the activation of rhodopsin, a G protein-coupled receptor (GPCR), and its functional interactions with G protein and arrestin. The protein reagents used for these studies come from native disc membranes or heterologous expression, and ...

journal_title：Methods in enzymology

authors： Sommer ME,Elgeti M,Hildebrand PW,Szczepek M,Hofmann KP,Scheerer P

更新日期：2015-01-01 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：:With the rapid development of nanotechnology, inorganic magnetic nanoparticles, especially iron oxide nanoparticles (IOs), have emerged as great vehicles for biomedical diagnostic and therapeutic applications. In order to rationally design IO-based gene delivery nanovectors, surface modification is essential and deter...

journal_title：Methods in enzymology

authors： Liu G,Wang Z,Lee S,Ai H,Chen X

更新日期：2012-01-01 00:00:00

abstract：:Mitochondrial RNAs in the myxomycete Physarum polycephalum differ from the templates from which they are transcribed in defined ways. Most transcripts contain nucleotides that are not present in their respective genes. These "extra" nucleotides are added during RNA synthesis by an unknown mechanism. Other differences ...

journal_title：Methods in enzymology

authors： Byrne EM,Visomirski-Robic L,Cheng YW,Rhee AC,Gott JM

更新日期：2007-01-01 00:00:00

abstract：:Tubulin is known to form high-affinity complexes with certain G proteins. The formation of such complexes allows tubulin to activate Galpha and fosters a system whereby elements of the cytoskeleton can influence G-protein signaling. This article describes the interaction between tubulin and G proteins and discusses me...

journal_title：Methods in enzymology

authors： Rasenick MM,Donati RJ,Popova JS,Yu JZ

更新日期：2004-01-01 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：:The hypoxia-inducible factor (HIF) is a heterodimeric basic helix-loop-helix (bHLH) transcription factor that controls the mammalian cellular transcriptional response to low oxygen tension by up-regulating genes including glycolytic enzymes and angiogenic factors, such as the vascular endothelial growth factor (VEGF)....

journal_title：Methods in enzymology

authors： Doedens A,Johnson RS

更新日期：2007-01-01 00:00:00

abstract：:The current treatment of neurological and psychiatric diseases is far beyond being satisfactory. In addition to highly complex disease mechanisms, the blood-brain barrier (BBB) also remains as a challenge by limiting the delivery of the majority of currently available therapeutics to the central nervous system. Severa...

journal_title：Methods in enzymology

authors： Yemişci M,Gürsoy-Özdemir Y,Caban S,Bodur E,Capan Y,Dalkara T

更新日期：2012-01-01 00:00:00

abstract：:As for other GPCRs, the oligomerization of glycoprotein hormone receptors (GPHRs) appears as critical event for receptor function. By means of modern techniques based on the BRET or FRET principle, GPHR oligomerization has been reported to explain several physiological and pathological conditions. In particular, the p...

journal_title：Methods in enzymology

authors： Bonomi M,Persani L

更新日期：2013-01-01 00:00:00

abstract：:The enzymes involved in mitochondrial oxidative phosphorylation (OXPHOS) are coassembled into higher ordered supercomplexes within the mitochondrial inner membrane. The cytochrome bc(1)-cytochrome c oxidase (COX) supercomplex is formed by the coassociation of the two electron transport chain complexes, the cytochrome ...

journal_title：Methods in enzymology

authors： Stuart RA

更新日期：2009-01-01 00:00:00

abstract：:We describe the basic principles for imaging frozen-hydrated specimens in a transmission electron microscope and provide a step-by-step guide to a new user, from starting up and aligning the microscope, to loading a cryo-grid into the specimen holder and inserting the holder into the microscope, to setting up the low ...

journal_title：Methods in enzymology

authors： Sun J,Li H

更新日期：2010-01-01 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：:Membrane proteins (MPs) are insoluble in aqueous media as a result of incompatibility between the hydrophilic property of the solvent molecules and the hydrophobic nature of MP surfaces, normally associated with lipid membranes. Amphipathic compounds are necessary for extraction of these macromolecules from the native...

journal_title：Methods in enzymology

authors： Sadaf A,Cho KH,Byrne B,Chae PS

更新日期：2015-01-01 00:00:00

abstract：:Polyacrylamide gel electrophoresis under native conditions (native PAGE) is a well-established and versatile method for probing nucleic acid conformation and nucleic acid-protein interactions. Native PAGE has been used to measure RNA folding equilibria and kinetics under a wide variety of conditions. Advantages of thi...

journal_title：Methods in enzymology

authors： Woodson SA,Koculi E

更新日期：2009-01-01 00:00:00

abstract：:Enzymology is approaching an era where many problems can benefit from computational studies. While ample challenges remain in quantitatively predicting behavior for many enzyme systems, the insights that often come from computations are an important asset for the enzymology community. Here we provide a primer for enzy...

journal_title：Methods in enzymology

authors： Roston D,Cui Q

更新日期：2016-01-01 00:00:00

abstract：:The functional properties of proteins are decided not only by their relatively rigid overall structures, but even more importantly, by their dynamic properties. In a protein, some regions of structure exhibit highly correlated or anti-correlated motions with others, some are highly dynamic but uncorrelated, while othe...

journal_title：Methods in enzymology

authors： Yu H,Dalby PA

更新日期：2020-01-01 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：: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：: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：: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：:Precisely imaging and tracking dynamic biological processes in live cells are crucial for both fundamental research in life sciences and biomedical applications. Nonfluorescent nanoparticles are emerging as important optical probes in live-cell imaging because of their excellent photostability, large optical cross sec...

journal_title：Methods in enzymology

authors： Wang G,Fang N

更新日期：2012-01-01 00:00:00

abstract：:Formation of 5-methyluridine (ribothymidine) at position 54 of the T-psi loop of tRNA is catalyzed by site-specific tRNA methyltransferases (tRNA[uracil-54,C5]-MTases). In eukaryotes and many bacteria, the methyl donor for this reaction is generally S-adenosyl-L-methionine (S-AdoMet). However, in other bacteria, like ...

journal_title：Methods in enzymology

authors： Urbonavicius J,Brochier-Armanet C,Skouloubris S,Myllykallio H,Grosjean H

更新日期：2007-01-01 00:00:00

abstract：:In principle, the long emission lifetimes of lanthanide chelates should enable their ultrasensitive detection in biological systems by time-resolved optical microscopy. However, most lanthanide-imaging systems cannot achieve sensitivities that exceed those of conventional fluorescence microscopes, since they are limit...

journal_title：Methods in enzymology

authors： Ciepla P,Cho U,Chen JK

更新日期：2020-01-01 00:00:00

abstract：:This chapter outlines the approaches and techniques that can be used to analyze the regulation of antibiotic production in streptomycetes. It describes how to isolate antibiotic nonproducing and overproducing mutants by UV, nitrosoguanidine (NTG), transposon, and insertion mutagenesis, and then how to use those mutant...

journal_title：Methods in enzymology

authors： Bibb M,Hesketh A

更新日期：2009-01-01 00:00:00

abstract：:Mutations in regulators and effectors of the Rho GTPases underlie various forms of mental retardation (MR). Among them, oligophrenin-1 (OPHN1), which encodes a Rho-GTPase activating protein, was one of the first Rho-linked MR genes identified. Upon characterization of OPHN1 in hippocampal brain slices, we obtained evi...

journal_title：Methods in enzymology

authors： Kasri NN,Govek EE,Van Aelst L

更新日期：2008-01-01 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