Receptor-driven identification of novel human A₃ adenosine receptor antagonists as potential therapeutic agents.

abstract：:Expressed protein ligation (EPL) combines two methods to ligate a synthetic peptide to a recombinant protein. Native chemical ligation (NCL) is a process in which two synthesized peptides are ligated by reaction of a C-terminal thioester on one peptide with an N-terminal cysteine residue of another protein. The chemis...

journal_title：Methods in enzymology

authors： Mitchell SF,Lorsch JR

更新日期：2014-01-01 00:00:00

abstract：:This chapter outlines methods that can be applied to determine the levels of lipids in cells and tissues. In particular, the methods focus upon the extraction and analysis of those lipids critical for monitoring signal transduction pathways. The methods address the analysis of the phosphoinositides, the lipid agonists...

journal_title：Methods in enzymology

authors： Wakelam MJ,Pettitt TR,Postle AD

更新日期：2007-01-01 00:00:00

abstract：:Amyloid aggregates are associated with a number of mammalian neurodegenerative diseases. Infectious aggregates of the mammalian prion protein PrP(sc) are hallmarks of transmissible spongiform encephalopathies in humans and cattle (Griffith, 1967; Legname et al., 2004; Prusiner, 1982; Silveira et al., 2004). Likewise, ...

journal_title：Methods in enzymology

authors： Bagriantsev SN,Kushnirov VV,Liebman SW

更新日期：2006-01-01 00:00:00

abstract：:Microtubules are dynamic polymers of the cytoskeleton, which play important roles in cell division, polarization, and intracellular transport. Self-assembly of microtubule polymer from αβ-tubulin heterodimers is highly variable, with stochastic switching between alternate states of net growth and net shortening, a phe...

journal_title：Methods in enzymology

authors： Prahl LS,Castle BT,Gardner MK,Odde DJ

更新日期：2014-01-01 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：:A genetic interaction occurs when the combination of two mutations leads to an unexpected phenotype. Screens for synthetic genetic interactions have been used extensively to identify genes whose products are functionally related. In particular, synthetic lethal genetic interactions often identify genes that buffer one...

journal_title：Methods in enzymology

authors： Baryshnikova A,Costanzo M,Dixon S,Vizeacoumar FJ,Myers CL,Andrews B,Boone C

更新日期：2010-01-01 00:00:00

abstract：:Epitope tagging has provided a useful experimental strategy with widespread applicability. The ample variety of epitope tag systems that have been put to use to date provide a collection of attributes relevant to virtually any experimental system. As a consequence, epitope tagging will continue to be a valuable tool f...

journal_title：Methods in enzymology

authors： Fritze CE,Anderson TR

更新日期：2000-01-01 00:00:00

abstract：:PqqE is the first enzyme in the biosynthetic pathway of the redox cofactor pyrroloquinoline quinone (PQQ), catalyzing the formation of a carbon-carbon bond in the precursor peptide PqqA. PqqE is a radical S-adenosyl-l-methionine (SAM) (RS) enzyme, a family of enzymes that use the reductive cleavage of a [4Fe-4S] clust...

journal_title：Methods in enzymology

authors： Zhu W,Martins AM,Klinman JP

更新日期：2018-01-01 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：: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：:Autophagy is a catabolic process that is crucial for cellular homeostasis and adaptive response to changing environments. Importantly, autophagy has been shown to be induced in many longevity-associated scenarios and to be required to maintain lifespan extension. Notably, autophagy is a highly conserved cellular proce...

journal_title：Methods in enzymology

authors： Kainz K,Tadic J,Zimmermann A,Pendl T,Carmona-Gutierrez D,Ruckenstuhl C,Eisenberg T,Madeo F

更新日期：2017-01-01 00:00:00

abstract：:Benzylisoquinoline alkaloids (BIAs) constitute a diverse class of plant secondary metabolites that includes the opiate analgesics morphine and codeine. Collectively, BIAs exhibit a myriad of pharmacological activities, including antimicrobial, antitussive, antispasmodic, and anticancer properties. Despite 2500 known B...

journal_title：Methods in enzymology

authors： Pyne ME,Narcross L,Fossati E,Bourgeois L,Burton E,Gold ND,Martin VJ

更新日期：2016-01-01 00:00:00

abstract：:Glass microfluidic devices have been fabricated to monitor the secretion of glycerol or fatty acids from cultured murine 3T3-L1 adipocytes. In the current studies, adipocytes are perfused in a reversibly sealed cell chamber, and secreted products are analyzed by enzyme assay on either a single- or dual-chip device. Th...

journal_title：Methods in enzymology

authors： Dugan CE,Kennedy RT

更新日期：2014-01-01 00:00:00

abstract：:Endosomes are isolated from rat liver using high-speed centrifugation through sucrose density gradients. They are distinguishable from Golgi elements, with which they coisolate, by their capacity to concentrate internalized protein ligands (viz., insulin and epidermal growth factor (EGF)) in receptor-bound intact form...

journal_title：Methods in enzymology

authors： Posner BI,Bergeron JJ

更新日期：2014-01-01 00:00:00

abstract：:Autophagy is an extremely dynamic process that mediates the rapid degradation of intracellular components in response to different stress conditions. The autophagic response is executed by specific protein complexes, whose function is regulated by posttranslational modifications and interactions with positive and nega...

journal_title：Methods in enzymology

authors： Antonioli M,Ciccosanti F,Dengjel J,Fimia GM

更新日期：2017-01-01 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：: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：:Cell-free (CF) protein expression has emerged as one of the most efficient production platforms for membrane proteins. Central bottlenecks prevalent in conventional cell-based expression systems such as mistargeting, inclusion body formation, degradation as well as product toxicity can be addressed by taking advantage...

journal_title：Methods in enzymology

authors： Henrich E,Dötsch V,Bernhard F

更新日期：2015-01-01 00:00:00

abstract：:A direct transport route from early/recycling endosome (EE/RE) to the trans-Golgi network (TGN) is exploited by Shiga toxin (Mallard et al., 1998) and TGN38 (Ghosh et al., 1998). To facilitate the study of this pathway, both in vivo and in vitro transport assays using recombinant Shiga toxin B fragments (STxB) as prot...

journal_title：Methods in enzymology

authors： Tai G,Lu L,Johannes L,Hong W

更新日期：2005-01-01 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：:The number of genetically modified mouse lines has been increasing exponentially in the past few decades. In order to safeguard them from accidental loss and genetic drifting, to reduce animal housing cost, and to efficiently distribute them around the world, it is important to cryopreserve these valuable genetic reso...

journal_title：Methods in enzymology

authors： Takahashi H,Liu C

更新日期：2010-01-01 00:00:00

abstract：:Isotopic enrichment of biomacromolecules is a widely used technique that enables the investigation of the structural and dynamic properties to provide information not accessible with natural abundance isotopic composition. This study reports an approach for deuterium incorporation into bacterial cellulose. A media for...

journal_title：Methods in enzymology

authors： O'Neill H,Shah R,Evans BR,He J,Pingali SV,Chundawat SP,Jones AD,Langan P,Davison BH,Urban V

更新日期：2015-01-01 00:00:00

abstract：:This study designed, synthesized, and characterized a number of new heterobifunctional photocleavable cross-linking reagents that may be used to photomodulate the activity of proteins or to prepare caged fluorescent dyes. Biomolecules or fluorophores caged via a thiol group with the BNBASE reagent can be covalently li...

journal_title：Methods in enzymology

authors： Marriott G,Ottl J

更新日期：1998-01-01 00:00:00

abstract：:Acoustic sensors probe the response of a thin layer to the mechanical displacement associated with an acoustic wave. Acoustic measurements provide two simultaneous time-resolved signals; one signal is related to the velocity or frequency of the acoustic wave and is mainly a function of adsorbed mass, while the second ...

journal_title：Methods in enzymology

authors： Melzak K,Tsortos A,Gizeli E

更新日期：2009-01-01 00:00:00

abstract：:Nonribosomal peptides are natural products typically of bacterial and fungal origin. These highly complex molecules display a broad spectrum of biological activities, and have been exploited for the development of immunosuppressant, antibiotic, anticancer, and other therapeutic agents. The nonribosomal peptides are as...

journal_title：Methods in enzymology

authors： Wilkinson B,Micklefield J

更新日期：2009-01-01 00:00:00

abstract：:The method presented here is intended as a compromise between finding a good overall alignment and the time taken to do so. Many multiple alignment algorithms spend an excessively large amount of effort trying to find the best global alignment. This time is often ill spent because the results of the standard dynamic p...

journal_title：Methods in enzymology

authors： Taylor WR

更新日期：1990-01-01 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：: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：:Crystallographic studies on flavodiiron proteins (FDPs) have revealed that the common sequence core ( approximately 400 residues) that defines this protein family comprises two structural domains. The N-terminal domain (of approximately 250 residues) displays a metallo-beta-lactamase-like-fold, being indeed structural...

journal_title：Methods in enzymology

authors： Vicente JB,Carrondo MA,Teixeira M,Frazão C

更新日期：2008-01-01 00:00:00

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