Comparative Sequence-Function Analysis of the Major Facilitator Superfamily: The "Mix-and-Match" Method.

abstract：:Liposomes show great promise as intravenous drug delivery vehicles, but it is often difficult to combine stability in the circulation with rapid, targeted release at the site of interest. Targeting to specific tissues requires developing highly specific ligands with strong affinities to receptors overexpressed on dise...

journal_title：Methods in enzymology

authors： Wu G,Mikhailovsky A,Khant HA,Zasadzinski JA

更新日期：2009-01-01 00:00:00

abstract：:Biotinylated molecules are extensively employed in bioanalytics and biotechnology. The currently available assays for quantification of biotin groups suffer from low sensitivity, low accuracy, or provide highly variable responses for different biotin derivatives. We developed a competitive binding assay in which avidi...

journal_title：Methods in enzymology

authors： Oberbichler E,Wiesauer M,Schlögl E,Stangl J,Faschinger F,Knör G,Gruber HJ,Hytönen VP

更新日期：2020-01-01 00:00:00

abstract：:The Rcs phosphorelay, consisting of a hybrid sensor kinase, a phosphotransferase, and a response regulator, regulates a large number of bacterial functions. These include capsule production, the target originally defined for these regulators, a small regulatory RNA, and a growing list of additional genes, many of unkn...

journal_title：Methods in enzymology

authors： Majdalani N,Gottesman S

更新日期：2007-01-01 00:00:00

abstract：:Focal radiation therapy has the potential to generate systemic tumor-targeting immune responses so potent as to eradicate anatomically distant, non-irradiated malignant lesions, a phenomenon commonly referred to as "the abscopal response." In cancer patients, bona fide abscopal responses are rare, although the recent ...

journal_title：Methods in enzymology

authors： Rodriguez-Ruiz ME,Yamazaki T,Buqué A,Bloy N,Silva VAO,Stafford L,Sato A,Galluzzi L

更新日期：2020-01-01 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：:Multiprotein machines drive virtually all primary cellular processes. Modular multidomain proteins are widely distributed within these dynamic complexes because they provide the flexibility needed to remodel structure as well as rapidly assemble and disassemble components of the machinery. Understanding the functional...

journal_title：Methods in enzymology

authors： Thompson MK,Ehlinger AC,Chazin WJ

更新日期：2017-01-01 00:00:00

abstract：:There are estimated to be >300,000 plant species, producing >200,000 metabolites. Many of these metabolites are restricted to specific plant lineages and are referred to as "specialized" metabolites. These serve varied functions in plants including defense against biotic and abiotic stresses, plant-plant and plant-mic...

journal_title：Methods in enzymology

authors： Fan P,Moghe GD,Last RL

更新日期：2016-01-01 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：:Embryonic stem (ES) cells are cells derived from the inner cell mass of a blastocyst stage embryo. These self-renewing multipotent cells are able to differentiate to the three embryonic germ layers, the endoderm, ectoderm, and mesoderm, and are thus able to produce virtually all cell types. The ES cell capacity to gen...

journal_title：Methods in enzymology

authors： Goldberg-Cohen I,Beck G,Ziskind A,Itskovitz-Eldor J

更新日期：2006-01-01 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：: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：:Of all arginine catabolic enzymes, the arginases and nitric oxide (NO) synthases are the ones that are of greatest interest to many investigators. Mammalian arginases catalyze the hydrolysis of arginine to ornithine and urea and are composed of two distinct isozymes: arginase I, located within the cytosol, and arginas...

journal_title：Methods in enzymology

authors： Kepka-Lenhart D,Ash DE,Morris SM

更新日期：2008-01-01 00:00:00

abstract：:Neurofilament biology is important to understanding structural properties of axons, such as establishment of axonal diameter by radial growth. In order to study the function of neurofilaments, a series of genetically modified mice have been generated. Here, we describe a brief history of genetic modifications used to ...

journal_title：Methods in enzymology

authors： Jones MR,Villalón E,Garcia ML

更新日期：2016-01-01 00:00:00

abstract：:The Gene Expression Omnibus (GEO) repository at the National Center for Biotechnology Information archives and freely distributes high-throughput molecular abundance data, predominantly gene expression data generated by DNA microarray technology. The database has a flexible design that can handle diverse styles of bot...

journal_title：Methods in enzymology

authors： Barrett T,Edgar R

更新日期：2006-01-01 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：:Mdm2 is a negative regulator of p53 activity and functions as an E3 ubiquitin ligase of p53. Inhibition of mdm2 E3 ligase activity will block ubiquitination and subsequent proteasome-mediated degradation of p53, resulting in the stabilization of p53 protein that could lead to the restoration of its tumor-suppressor ac...

journal_title：Methods in enzymology

authors： Auger KR,Copeland RA,Lai Z

更新日期：2005-01-01 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：:Nanotechnology offers a new platform for therapeutic delivery of antiretrovirals to the central nervous system (CNS) where human immunodeficiency virus (HIV-1) is sequestered in patients with HIV-1-associated neurological disorders (HAND). HAND is a spectrum of neurocognitive disorders that continue to persist in HIV-...

journal_title：Methods in enzymology

authors： Mahajan SD,Law WC,Aalinkeel R,Reynolds J,Nair BB,Yong KT,Roy I,Prasad PN,Schwartz SA

更新日期：2012-01-01 00:00:00

abstract：:The gasdermin (GSDM) family consists of gasdermin A (GSDMA), B (GSDMB), C (GSDMC), D (GSDMD), E or DNFA5 (GSDME), and DFNB59 in human. Expressed in the skin, gastrointestinal tract, and various immune cells, GSDMs mediate homeostasis and inflammation upon activation by caspases and unknown proteases. In particular, GS...

journal_title：Methods in enzymology

authors： Xia S,Ruan J,Wu H

更新日期：2019-01-01 00:00:00

abstract：:Advanced glycation is one of the major pathophysiological posttranslational modifications. Under hyperglycemic conditions or oxidative stress, proteins and DNA are nonenzymatically modified by oxidative glycation and converted to advanced glycation endproducts (AGEs), which induce the loss of protein functions or apop...

journal_title：Methods in enzymology

authors： Inagi R

更新日期：2011-01-01 00:00:00

abstract：:Biominerals have complex and heterogeneous architectures, hence diffraction experiments with spatial resolutions between 500 nm and 10 μm are extremely useful to characterize them. X-ray beams in this size range are now routinely produced at many synchrotrons. This chapter provides a review of the different hard X-ray...

journal_title：Methods in enzymology

authors： Tamura N,Gilbert PU

更新日期：2013-01-01 00:00:00

abstract：:Aliphatic carbon-halogen (C-X) bonds are prevalent in modern pharmaceuticals and bioactive natural products. Three distinct chemical strategies are known in Nature to generate these structural motifs. The first is via the nucleophilic substitution at a prefunctionalized electrophilic carbon center with a halide anion ...

journal_title：Methods in enzymology

authors： Liu X

更新日期：2018-01-01 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：:The human cannabinoid receptor, CB1, has been difficult to purify in a functional form, hampering structural and biophysical studies. Here, we present our approaches for obtaining pure, detergent solubilized, functional CB1. We also discuss our site-directed fluorescence labeling (SDFL) methods for identifying differe...

journal_title：Methods in enzymology

authors： Fay JF,Farrens DL

更新日期：2017-01-01 00:00:00

abstract：:Reviewing the circumstances that have led to the first monoclonal antibody against the disease-associated form of PrP, we consider the availability of PrP knockout mice and recombinant PrP, as well as a reliable conformational screening protocol as being important prerequisites for a successful immunization approach. ...

journal_title：Methods in enzymology

authors： Korth C,Streit P,Oesch B

更新日期：1999-01-01 00:00:00

abstract：:Fluorescence-based imaging regimes require exposure of living samples under study to high intensities of focused incident illumination. An often underestimated, overlooked, or simply ignored fact in the design of any experimental imaging protocol is that exposure of the specimen to these excitation light sources must ...

journal_title：Methods in enzymology

authors： Tinevez JY,Dragavon J,Baba-Aissa L,Roux P,Perret E,Canivet A,Galy V,Shorte S

更新日期：2012-01-01 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：:Physical, chemical, and biological processes commonly discriminate among stable isotopes. Therefore, the stable isotope compositions of biomass, growth substrates, and products often carry the isotopic fingerprints of the processes that shape them. Therefore, measuring isotope fractionation by enzymes and cultures of ...

journal_title：Methods in enzymology

authors： Scott KM,Fox G,Girguis PR

更新日期：2011-01-01 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：:Sedimentation velocity experiments measure the transport of molecules in solution under centrifugal force. Here, we describe a method for monitoring the sedimentation of very large biological molecular assemblies using the interference optical systems of the analytical ultracentrifuge. The mass, partial-specific volum...

journal_title：Methods in enzymology

authors： Rogge RA,Hansen JC

更新日期：2015-01-01 00:00:00