Droplet confinement and fluorescence measurement of single molecules.

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

abstract：:Scanning fluorescence correlation spectroscopy (sFCS) is the generic term for a group of fluorescence correlation techniques where the measurement volume is moved across the sample in a defined way. The introduction of scanning is motivated by its ability to alleviate or remove several distinct problems often encounte...

journal_title：Methods in enzymology

authors： Petrásek Z,Ries J,Schwille P

更新日期：2010-01-01 00:00:00

abstract：:The history of free radical biochemistry is briefly reviewed in respect to major trend shifts from the focus on radiation damage toward enzymology of radical production and removal and ultimately the role of radicals, hydroperoxides, and related fast reacting compounds in metabolic regulation. Selected aspects of the ...

journal_title：Methods in enzymology

authors： Flohé L

更新日期：2010-01-01 00:00:00

abstract：:Throughout the history of single-particle electron microscopy (EM), automated technologies have seen varying degrees of emphasis and development, usually depending upon the contemporary demands of the field. We are currently faced with increasingly sophisticated devices for specimen preparation, vast increases in the ...

journal_title：Methods in enzymology

authors： Lyumkis D,Moeller A,Cheng A,Herold A,Hou E,Irving C,Jacovetty EL,Lau PW,Mulder AM,Pulokas J,Quispe JD,Voss NR,Potter CS,Carragher B

更新日期：2010-01-01 00:00:00

abstract：:Galectins are a family of animal lectins with affinity for beta-galactosides. By using recombinant proteins, a number of galectins have been shown to interact with cell-surface and extracellular matrix glycoconjugates through lectin-carbohydrate interactions. Through this action, they can affect a variety of cellular ...

journal_title：Methods in enzymology

authors： Hsu DK,Yang RY,Liu FT

更新日期：2006-01-01 00:00:00

abstract：:Site-specific protein labeling can be used to monitor protein motions and interactions in real time using Förster resonance energy transfer (FRET). While there are many fluorophores available for protein labeling, few FRET pairs are suitable for monitoring intramolecular protein motions without being disruptive to pro...

journal_title：Methods in enzymology

authors： Jones CM,Venkatesh Y,Petersson EJ

更新日期：2020-01-01 00:00:00

abstract：:Chromatin plays a vital role in regulating cellular processes that occur on the DNA. Modulation of chromatin structure is conducted through interactions with binding factors that direct critical actions such as posttranslational modifications, nucleosome remodeling, and incorporation of histone variants. Specific fact...

journal_title：Methods in enzymology

authors： Winkler DD,Luger K,Hieb AR

更新日期：2012-01-01 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：:The protein kinase C (PKC) family of isoenzymes may be a crucial player in transducing H2O2-induced signaling in a wide variety of physiological and pathophysiological processes. PKCs contain unique structural features that make them highly susceptible to oxidative modification. Depending on the site of oxidation and ...

journal_title：Methods in enzymology

authors： Gopalakrishna R,McNeill TH,Elhiani AA,Gundimeda U

更新日期：2013-01-01 00:00:00

abstract：:There is currently no known genetic disease linked to prolactin (PRL) or its receptor (PRLR) in humans. Recently, we identified three missense variants of the PRLR in patients presenting with breast tumors. Two of them (named PRLR(I146L) and PRLR(I76V)) had been reported earlier, but failed to draw much attention beca...

journal_title：Methods in enzymology

authors： Goffin V,Bogorad RL,Touraine P

更新日期：2010-01-01 00:00:00

abstract：:In vitro assembly of eukaryotic translation initiation complexes requires purification of ribosomal subunits, eukaryotic initiation factors, and initiator tRNA from natural sources and therefore yields only limited material for functional and structural studies. In this chapter, we describe a robust, affinity chromato...

journal_title：Methods in enzymology

authors： Locker N,Lukavsky PJ

更新日期：2007-01-01 00:00:00

abstract：:The RNA degradosome is a bacterial protein machine devoted to RNA degradation and processing. In Escherichia coli, it is typically composed of the endoribonuclease RNase E, which also serves as a scaffold for the other components: the exoribonuclease PNPase, the RNA helicase RhlB, and enolase. The variable presence of...

journal_title：Methods in enzymology

authors： Mauri P,Dehò G

更新日期：2008-01-01 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：:By far the highest resolution of all separation techniques for intact proteins in a single analytical run continues to be by the combination of isoelectric focusing (IEF) and SDS-PAGE, originally introduced by O'Farrell [(1975). J. Biol. Chem.250, 4007-4021]. This analytical platform has seen a number of significant a...

journal_title：Methods in enzymology

authors： Friedman DB,Hoving S,Westermeier R

更新日期：2009-01-01 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：:Fluorescence live-cell RNA imaging to monitor the intracellular localization and dynamics of the target RNA is a challenging subject. One of the difficulties to achieve this is to establish a precise method to enable a fluorescent labeling to the target RNA in living cells. Technologies to reduce the background fluore...

journal_title：Methods in enzymology

authors： Yoshimura H,Ozawa T

更新日期：2016-01-01 00:00:00

abstract：:To elucidate the molecular mechanisms of vertebrate color vision, it is essential to establish associations between amino acid substitutions and the directions of lambda max shifts of visual pigments. In this way, we can identify critical amino acid changes that may be responsible for lambda max shifts of visual pigme...

journal_title：Methods in enzymology

authors： Yokoyama S

更新日期：2000-01-01 00:00:00

abstract：:Essentially the same steps are required to solve the crystal structure of a serpin as for any other protein: produce and purify protein, grow crystals, collect diffraction data, find estimates of the phase angles, and then refine and validate the structure. For the phasing step, experimental phasing methods involving ...

journal_title：Methods in enzymology

authors： Read RJ,Zhou A,Stein PE

更新日期：2011-01-01 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：: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：:Use of fluorescence in biology and biochemistry for imaging and characterizing equilibrium and dynamic processes is growing exponentially. Much progress has been made in the last few years on the microscopic understanding of the underlying principles of what controls the wavelength and quenching of fluorescence in bio...

journal_title：Methods in enzymology

authors： Callis PR

更新日期：2011-01-01 00:00:00

abstract：:A fundamental concept of phosphorylation-mediated signaling is the precise switching between discrete functional conformations. According to the traditional view, phosphorylation induces a new, active conformation. In this chapter, a series of NMR experiments performed on a response regulator are described that challe...

journal_title：Methods in enzymology

authors： Gardino AK,Kern D

更新日期：2007-01-01 00:00:00

abstract：:Contemporary high-throughput sequencing methods, notably RNA-Seq, permit the systematic identification and characterization of transcripts whose levels change significantly in response to altered biological states. We have described methods for the application of this methodology to a definition of the transcripts reg...

journal_title：Methods in enzymology

authors： He F,Celik A,Baker R,Jacobson A

更新日期：2018-01-01 00:00:00

abstract：:Model building studies may be used to supplement structurally low resolution experimental data with detailed three-dimensional hypothetical atomic models. Because of the strong relation between structure and function in biological molecules such models may give a consistent, integral view of a wealth of experimental d...

journal_title：Methods in enzymology

authors： von Kitzing E

更新日期：1992-01-01 00:00:00

abstract：:Two factors, climate change brought on by rising atmospheric CO2 levels and the accelerating shift toward renewable energy sources, have together worked to heighten interest in understanding how biological catalysts so effectively bring about the reduction of CO2 to formate, with potential applications for both biorem...

journal_title：Methods in enzymology

authors： Niks D,Hille R

更新日期：2018-01-01 00:00:00

abstract：:The three-dimensional (3D) organization of the genome is important for chromatin regulation. This organization is nonrandom and appears to be tightly correlated with or regulated by chromatin state and scaffolding proteins. To understand how specific DNA and chromatin elements contribute to the functional organization...

journal_title：Methods in enzymology

authors： Harr JC,Reddy KL

更新日期：2016-01-01 00:00:00

abstract：:Guanine nucleotide exchange factors (GEFs) are essential regulators of the spatiotemporal conditions of small GTP-binding protein (SMG) activation. Their cellular activities combine the biochemical stimulation of GDP/GTP exchange, which leads to the active conformation of the SMG, to the detection of upstream signals ...

journal_title：Methods in enzymology

authors： Zeeh JC,Antonny B,Cherfils J,Zeghouf M

更新日期：2008-01-01 00:00:00

abstract：:Complex synthetic schemes catalyzed by multienzyme systems immobilized on solid materials are gaining momentum in chemical biomanufacturing. These systems harness the high chemo-, regio-, and stereoselectivity of the enzymes and the recyclability of the heterogeneous catalysts. Moreover, when the enzymes become part o...

journal_title：Methods in enzymology

authors： López-Gallego F

更新日期：2019-01-01 00:00:00

abstract：:Phospholipase A(2)s (PLA(2)s) are a superfamily of enzymes involved in production of a wide variety of lipid mediators, including arachidonic acid, lysophospholipids, platelet activation factor, and eicosanoids. Fluorescence-based, real-time cellular activity assays for PLA(2)s have been developed as a tool for studyi...

journal_title：Methods in enzymology

authors： Manna D,Cho W

更新日期：2007-01-01 00:00:00

abstract：:The JNKs (c-Jun N-terminal protein kinases) play important roles in a variety of physiological and pathological functions including induction of apoptosis. A major pathway by which JNKs are activated in response to apoptotic stimuli includes the GTP*Rac1-binding scaffold protein POSH (plenty of SH3s). POSH acts as a s...

journal_title：Methods in enzymology

authors： Xu Z,Greene LA

更新日期：2006-01-01 00:00:00