Characterization of oligophrenin-1, a RhoGAP lost in patients affected with mental retardation: lentiviral injection in organotypic brain slice cultures.

abstract：:Ubiquitin (Ub)-mediated protein degradation is a key cellular defense mechanism that detects and eliminates defective proteins. A major intracellular site of protein quality control degradation is the endoplasmic reticulum (ER), hence the term ER-associated degradation, or endoplasmic reticulum-associated degradation ...

journal_title：Methods in enzymology

authors： Mashahreh B,Reiss Y,Wiener R,Ravid T

更新日期：2019-01-01 00:00:00

abstract：:Perhaps the most important contribution of FLN is that it provides an experimental approach to relate physical changes in the protein to predicted dynamical behavior. It is clear that the sample is inhomogeneously broadened in a continuous manner, consistent with the damped motion of proteins. At the same time configu...

journal_title：Methods in enzymology

authors： Vanderkooi JM,Angiolillo PJ,Laberge M

更新日期：1997-01-01 00:00:00

abstract：:Rac binds tightly to lipid membranes through a lipid modification. The influence of the lipid membrane environment on the multiple interactions of Rac has not been well documented. In this chapter, we detail a method to prepare geranyl-geranylated Rac bound to liposomes of defined composition. With this method, one ca...

journal_title：Methods in enzymology

authors： Mesmin B,Antonny B

更新日期：2006-01-01 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：:G-protein-coupled receptors (GPCRs) form a superfamily of membrane proteins that play a crucial role in mediating physiological processes as well as pathogenesis of many critical diseases. They are one of the most successful drug targets, accounting for more than 30% of prescription drugs on the market today. Three-di...

journal_title：Methods in enzymology

authors： Vaidehi N,Pease JE,Horuk R

更新日期：2009-01-01 00:00:00

abstract：:Concomitant with the rapid development in biomedical knowledge, including the methods of molecular biology and proteomics, and the manufacture of ever more precise optical instruments, powerful lasers, and sophisticated microcomputing hardware and software, laser microdissection systems have emerged which are now ente...

journal_title：Methods in enzymology

authors： Willenberg HS,Walters R,Bornstein SR

更新日期：2002-01-01 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：: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：: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：:All cells in nature are covered with a dense and complex array of glycans. The total glycan repertoire expressed on cells, "the cellular glycome," varies at every level of biological organization, and in response to intrinsic and extrinsic stimuli. The cellular glycome is often referred to as the "cell face," which re...

journal_title：Methods in enzymology

authors： Tateno H,Kuno A,Itakura Y,Hirabayashi J

更新日期：2010-01-01 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：:Measurement of catalysis by MnSOD using direct observation of the UV absorbance of superoxide allows determination of steady-state catalytic constants. Stabilizing superoxide in aprotic solvents such as dimethyl sulfoxide permits the use of stopped-flow spectrophotometry, although significant information is lost in th...

journal_title：Methods in enzymology

authors： Silverman DN,Nick HS

更新日期：2002-01-01 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：:Hybrid proteins or chimeras are generated by recombination of protein fragments. In the course of evolution, this mechanism has led to major diversification of protein folds and their functionalities. Similarly, protein engineers have taken advantage of this attractive strategy to build new proteins. Methods that use ...

journal_title：Methods in enzymology

authors： Rico JA,Höcker B

更新日期：2013-01-01 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：:Delivery of drugs and molecules to eukaryote animal and plant cells can be enhanced when mediated by mesoporous silica nanoparticle (MSN) materials. This chapter describes, in detail, the synthesis of several different MSN materials used for the transport of various hydrophilic and hydrophobic drug molecules, neurotra...

journal_title：Methods in enzymology

authors： Fang IJ,Trewyn BG

更新日期：2012-01-01 00:00:00

abstract：:Mathematical computer-assisted simulations of episodic hormone, substrate, or metabolite release can be accomplished using explicit algebra and high-speed microprocessors. Such simulations are useful to describe the behavior of single or multiple pulse generators, as well as the expected output of these systems. Simul...

journal_title：Methods in enzymology

authors： Veldhuis JD,Johnson ML

更新日期：1994-01-01 00:00:00

abstract：:Considering that HIV-1 accumulates and replicates actively within lymphoid tissues, any strategy that will decrease viral stores in these tissues might be beneficial to the infected host. Follicular dendritic cells (FDC), B lymphocytes, antigen-presenting cells like macrophages, and activated CD4(+) T cells are abunda...

journal_title：Methods in enzymology

authors： Désormeaux A,Bergeron MG

更新日期：2005-01-01 00:00:00

abstract：:The combined use of rotating-frame relaxation methods, temperature-dependent measurements of line shapes and magnetization transfer experiments allows in favorable cases the examination in some detail of exchange processes that occur on the millisecond time scale. It is possible to determine not only the rate constant...

journal_title：Methods in enzymology

authors： Lane AN,Lefèvre JF

更新日期：1994-01-01 00:00:00

abstract：:Reliable modeling of RNA tertiary structures is key to both understanding these structures' roles in complex biological machines and to eventually facilitating their design for molecular computing and robotics. In recent years, a concerted effort to improve computational prediction of RNA structure through the RNA-Puz...

journal_title：Methods in enzymology

authors： Cheng CY,Chou FC,Das R

更新日期：2015-01-01 00:00:00

abstract：:We have used a shortened construct form of the CYP26A1 gene promoter, in a promoter-less vector with either luciferase (known as E4) or a red fluorescent protein, RFP (known as E4.2) as the reporter gene and examined their responses to retinoids in transfected HepG2 and HEK293T cells. The promoter responded linearly t...

journal_title：Methods in enzymology

authors： Zolfaghari R,Mattie FJ,Wei CH,Chisholm DR,Whiting A,Ross AC

更新日期：2020-01-01 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：:Antimicrosomal antibodies in inflammatory liver diseases all seem to be directed against members of the cytochrome P450 family of proteins. These autoantigens seem to be genetically polymorphic, the autoantibodies are inhibitory, and the autoepitopes are generally conserved among species. Anti-P450 autoantibodies shar...

journal_title：Methods in enzymology

authors： Manns MP,Johnson EF

更新日期：1991-01-01 00:00:00

abstract：:Peptides are multidentate chiral ligands capable of coordinating different metal ions. Nowadays, they can be obtained with high yield and purity, thanks to the advances on peptide/protein chemistry as well as in equipment (peptide synthesizers). Based on the identity and length of their amino acid sequences, peptides ...

journal_title：Methods in enzymology

authors： Lima LM,Iranzo O

更新日期：2016-01-01 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：: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：: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 epitope tagging approach offers advantages of economy, universality, and precision over the use of antibodies raised directly against a protein of interest. The latter strategy promises a potentially greater diversity of reagents and obviates the need to modify the protein, but it may not yield sufficiently high-a...

journal_title：Methods in enzymology

authors： Kolodziej PA,Young RA

更新日期：1991-01-01 00:00:00

abstract：:In the near future, computational tools and methods based on the mathematical modeling of biomedically relevant networks and pathways will be necessary for the design of therapeutic strategies that fight complex multifactorial diseases. Beyond the use of pharmacokinetic and pharmacodynamic approaches, we propose here ...

journal_title：Methods in enzymology

authors： Marin-Sanguino A,Gupta SK,Voit EO,Vera J

更新日期：2011-01-01 00:00:00

abstract：:This protocol describes the expression and analysis of membrane proteins produced in yeast, as illustrated with Yarrowia lipolytica and Pichia pastoris. Step by step, we explain how to generate a yeast strain expressing the membrane protein of interest, how to prepare a membrane protein sample from yeast, and how to a...

journal_title：Methods in enzymology

authors： Claes K,Guerfal M,Callewaert N

更新日期：2015-01-01 00:00:00