当前位置： SCI文献检索 > BIOCHEMISTRY期刊下所有文献 > Signal amplification between Gbetagamma release and PI3Kgamma-mediated PI(3,4,5)P3 formation monitored by a fluorescent Gbetagamma biosensor protein and repetitive two component total internal reflection/fluorescence redistribution after photobleaching an

Signal amplification between Gbetagamma release and PI3Kgamma-mediated PI(3,4,5)P3 formation monitored by a fluorescent Gbetagamma biosensor protein and repetitive two component total internal reflection/fluorescence redistribution after photobleaching an

Abstract：

:Phosphoinositide 3-kinase gamma (PI3Kgamma) is activated by Gbetagamma release after stimulation of Galpha i -coupled receptors, involving a recruitment of the enzyme to the plasma membrane via interaction of the regulatory subunit p101 or p87 with Gbetagamma. The receptor-mediated release of Gbetagamma was, however, insufficient to elicit a translocation of p101 observable by classical fluorescence microscopy approaches. Since the mobilities of plasma membrane-associated and cytosolic proteins differ strongly, small changes in the amount of plasma membrane association should be detectable by an altered diffusional behavior. Here, changes in mobility were monitored by fluorescence redistribution after photobleaching (FRAP) which was repetitively applied before and after stimulation of cells. To combine the advantages of total internal reflection (TIR) illumination, which preferentially excites fluorophors located at or near the plasma membrane, with that provided by the mobility information, we developed a combined TIR/FRAP setup which enabled us to point bleach parts of an image that was observed under TIR illumination. For FRAP data analysis, we introduce a convolution-based method and a global two component model. Using this TIR/FRAP approach, an increased plasma membrane association of the fluorescent Gbetagamma-binding domain of p101 after Gbetagamma release by G protein-coupled receptor stimulation could be detected and quantified. By comparing the translocation efficiency of this domain with that of YFP-GRP1(PH), a biosensor for the PI3Kgamma product PI(3,4,5)P3, we evaluate the signal amplification between Gbetagamma release and PI(3,4,5)P3 formation after activation of Galpha i -coupled receptors.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Tannert A,Voigt P,Burgold S,Tannert S,Schaefer M

doi

10.1021/bi800596b

subject

Has Abstract

pub_date

2008-10-28 00:00:00

pages

11239-50

issue

43

eissn

0006-2960

issn

1520-4995

journal_volume

47

pub_type

杂志文章

相关文献

BIOCHEMISTRY文献大全
  • Fluorescence correlation spectroscopy detects galanin receptor diversity on insulinoma cells.

    abstract：:Fluorescence correlation spectroscopy (FCS) allows the study of interactions of fluorescently labeled ligand with receptors in living cells at single-molecule detection sensitivity. From the autocorrelation functions of fluorescence intensity fluctuations, the diffusion time of molecules through the confocal volume is...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi010514q

    authors： Pramanik A,Olsson M,Langel U,Bartfai T,Rigler R

    更新日期：2001-09-11 00:00:00

  • Procatalytic ligand strain. Ionization and perturbation of 8-nitroxanthine at the urate oxidase active site.

    abstract：:The binding of the inhibitor 8-nitroxanthine to urate oxidase has been investigated by Raman and UV-visible absorption spectroscopy. The absorption maximum of 8-nitroxanthine shifts from 380 to 400 nm upon binding to the enzyme, demonstrating that the electronic structure of the ligand is perturbed. It has been propos...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi0507837

    authors： Doll C,Bell AF,Power N,Tonge PJ,Tipton PA

    更新日期：2005-08-30 00:00:00

  • Catalysis at the interface: the anatomy of a conformational change in a triglyceride lipase.

    abstract：:The crystal structure of an extracellular triglyceride lipase (from a fungus Rhizomucor miehei) inhibited irreversibly by diethyl p-nitrophenyl phosphate (E600) was solved by X-ray crystallographic methods and refined to a resolution of 2.65 A. The crystals are isomorphous with those of n-hexylphosphonate ethyl ester/...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00120a034

    authors： Derewenda U,Brzozowski AM,Lawson DM,Derewenda ZS

    更新日期：1992-02-11 00:00:00

  • Binding of O(2) and its reduction are both retarded by replacement of valine 279 by isoleucine in cytochrome c oxidase from Paracoccus denitrificans.

    abstract：:The crystal structure of the heme-copper oxidases suggested a putative channel of oxygen entry into the heme-copper site of O(2) reduction. Changing a conserved valine near this center in cytochrome bo(3) of Escherichia coli to isoleucine caused a significant increase in the apparent K(M) for oxygen with little or no ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi000123w

    authors： Riistama S,Puustinen A,Verkhovsky MI,Morgan JE,Wikström M

    更新日期：2000-05-30 00:00:00

  • Site-specific variations in RNA folding thermodynamics visualized by 2-aminopurine fluorescence.

    abstract：:The fluorescent base analogue 2-aminopurine (2-AP) is commonly used to study specific conformational and protein binding events involving nucleic acids. Here, combinations of steady-state and time-resolved fluorescence spectroscopy of 2-AP were employed to monitor conformational transitions within a model hairpin RNA ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi7011977

    authors： Ballin JD,Bharill S,Fialcowitz-White EJ,Gryczynski I,Gryczynski Z,Wilson GM

    更新日期：2007-12-11 00:00:00

  • Synergistic Biomineralization Phenomena Created by a Combinatorial Nacre Protein Model System.

    abstract：:In the nacre or aragonite layer of the mollusk shell, proteomes that regulate both the early stages of nucleation and nano-to-mesoscale assembly of nacre tablets from mineral nanoparticle precursors exist. Several approaches have been developed to understand protein-associated mechanisms of nacre formation, yet we sti...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/acs.biochem.6b00163

    authors： Chang EP,Roncal-Herrero T,Morgan T,Dunn KE,Rao A,Kunitake JA,Lui S,Bilton M,Estroff LA,Kröger R,Johnson S,Cölfen H,Evans JS

    更新日期：2016-04-26 00:00:00

  • Role of catalytic residues in enzymatic mechanisms of homologous ketosteroid isomerases.

    abstract：:Ketosteroid isomerase (KSI) is one of the most proficient enzymes catalyzing an allylic isomerization reaction at a diffusion-controlled rate. In this study of KSI, we have detailed the structures of its active site, the role of various catalytic residues, and have explained the origin of the its fast reactivity by ca...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi001629h

    authors： Oh KS,Cha SS,Kim DH,Cho HS,Ha NC,Choi G,Lee JY,Tarakeshwar P,Son HS,Choi KY,Oh BH,Kim KS

    更新日期：2000-11-14 00:00:00

  • Completely buried, non-ion-paired glutamic acid contributes favorably to the conformational stability of pyrrolidone carboxyl peptidases from hyperthermophiles.

    abstract：:Pyrrolidone carboxyl peptidases (PCPs) from hyperthermophiles have a structurally conserved and completely buried Glu192 in the hydrophobic core; in contrast, the corresponding residue in the mesophile protein is a hydrophobic residue, Ile. Does the buried ionizable residue contribute to stabilization or destabilizati...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi052610n

    authors： Kaushik JK,Iimura S,Ogasahara K,Yamagata Y,Segawa S,Yutani K

    更新日期：2006-06-13 00:00:00

  • A phosphotyrosyl mimetic peptide reverses impairment of insulin-stimulated translocation of GLUT4 caused by overexpression of PTP1B in rat adipose cells.

    abstract：:Biological actions of insulin are initiated by activation of the insulin receptor tyrosine kinase. Protein tyrosine phosphatases (PTPases) PTP1B and PTPalpha are known to dephosphorylate the insulin receptor and may contribute to insulin resistance in diseases such as diabetes. We previously reported that overexpressi...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi9816103

    authors： Chen H,Cong LN,Li Y,Yao ZJ,Wu L,Zhang ZY,Burke TR Jr,Quon MJ

    更新日期：1999-01-05 00:00:00

  • Characterizing the response of calcium signal transducers to generated calcium transients.

    abstract：:Cellular Ca2+ transients and Ca2+-binding proteins regulate physiological phenomena as diverse as muscle contraction, neurosecretion, and cell division. When Ca2+ is rapidly mixed with slow Ca2+ chelators, EGTA, or Mg2+/EDTA, artificial Ca2+ transients (ACTs) of varying duration (0.1-50 ms half-widths (hws)) and ampli...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi982495z

    authors： Davis JP,Tikunova SB,Walsh MP,Johnson JD

    更新日期：1999-03-30 00:00:00

  • Novel mechanism for the activation of rhodopsin kinase: implications for other G protein-coupled receptor kinases (GRK's).

    abstract：:ATP, its nonhydrolyzable analogue, AMP-PNP, and albumin were found to promote the dissociation of rhodopsin kinase from rod outer segments (ROS) containing photoactivated-rhodopsin (Rho*). These features were embodied in a protocol for the recovery of rhodopsin kinase from incubations containing ROS which had been sub...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi952480q

    authors： Dean KR,Akhtar M

    更新日期：1996-05-14 00:00:00

  • Identification of the cyclosporin-binding site in P-glycoprotein.

    abstract：:The binding site of cyclosporin A to P-glycoprotein was characterized by using a multidrug-resistant Chinese hamster ovary cell line. P-glycoprotein photolabeled with diazirine-cyclosporin A analogue was purified by a two-step process involving continuous elution electrophoresis followed by wheat germ agglutinin-agaro...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi981992c

    authors： Demeule M,Laplante A,Murphy GF,Wenger RM,Béliveau R

    更新日期：1998-12-22 00:00:00

  • Characterization of human glucocorticoid receptor complexes formed with DNA fragments containing or lacking glucocorticoid response elements.

    abstract：:Sucrose density gradient shift assays were used to study the interactions of human glucocorticoid receptors (GR) with small DNA fragments either containing or lacking glucocorticoid response element (GRE) DNA consensus sequences. When crude cytoplasmic extracts containing [3H]triamcinolone acetonide [( 3H]TA) labeled ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00431a003

    authors： Tully DB,Cidlowski JA

    更新日期：1989-03-07 00:00:00

  • The stereoselectivity and catalytic properties of Xanthobacter autotrophicus 2-[(R)-2-Hydroxypropylthio]ethanesulfonate dehydrogenase are controlled by interactions between C-terminal arginine residues and the sulfonate of coenzyme M.

    abstract：:2-[(R)-2-Hydroxypropylthio]ethanesulfonate (R-HPC) dehydrogenase (DH) catalyzes the reversible oxidation of R-HPC to 2-(2-ketopropylthio)ethanesulfonate (2-KPC) in a key reaction in the bacterial conversion of chiral epoxides to beta-keto acids. R-HPCDH is highly specific for the R-enantiomer of HPC, while a separate ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi049783h

    authors： Clark DD,Boyd JM,Ensign SA

    更新日期：2004-06-01 00:00:00

  • Chiral sulfoxidations catalyzed by rat liver cytochromes P-450.

    abstract：:The chirality of sulfoxidation catalyzed by two cytochrome P-450 isozymes purified from phenobarbital-induced rat liver was studied by using 4-tolyl ethyl sulfide as a substrate. Both P-450 isoenzymes, termed PB-1 and PB-4, when reconstituted with purified rat liver NADPH-cytochrome P-450 reductase and cytochrome b5, ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00539a032

    authors： Waxman DJ,Light DR,Walsh C

    更新日期：1982-05-11 00:00:00

  • A kinetic study of interactions of (Rp)- and (Sp)-adenosine cyclic 3',5'-phosphorothioates with type II bovine cardiac muscle adenosine cyclic 3',5'-phosphate dependent protein kinase.

    abstract：:The stereoselectivity of the adenosine cyclic 3',5'-phosphate (cAMP) binding sites on the regulatory subunit of the type II bovine cardiac muscle cAMP-dependent protein kinase was investigated by examining the interactions of (Rp)- and (Sp)-adenosine cyclic 3',5'-phosphorothioates (cAMPS) with these sites. While activ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00261a028

    authors： O'Brian CA,Roczniak SO,Bramson HN,Baraniak J,Stec WJ,Kaiser ET

    更新日期：1982-08-31 00:00:00

  • Kinetic unfolding mechanism of the inducible nitric oxide synthase oxygenase domain determined by time-resolved electrospray mass spectrometry.

    abstract：:The inducible nitric oxide synthase core oxygen domain (iNOS(COD)) is a homodimeric protein complex of ca. 100 kDa. In this work, the subunit disassembly and unfolding of the protein following a pH jump from 7.5 to 2.8 were monitored by on-line rapid mixing in conjunction with electrospray (ESI) time-of-flight mass sp...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi047684y

    authors： Wilson DJ,Rafferty SP,Konermann L

    更新日期：2005-02-22 00:00:00

  • The dual-specific active site of 7,8-diaminopelargonic acid synthase and the effect of the R391A mutation.

    abstract：:7,8-diaminopelargonic acid (DAPA) synthase (EC 2.6.1.62) is a pyridoxal phosphate (PLP)-dependent transaminase that catalyzes the transfer of the alpha-amino group from S-adenosyl-L-methionine (SAM) to 7-keto-8-aminopelargonic acid (KAPA) to form DAPA in the antepenultimate step in the biosynthesis of biotin. The wild...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi026339a

    authors： Eliot AC,Sandmark J,Schneider G,Kirsch JF

    更新日期：2002-10-22 00:00:00

  • Access to the active site of periplasmic nitrate reductase: insights from site-directed mutagenesis and zinc inhibition studies.

    abstract：:The periplasmic nitrate reductase (NapAB), a member of the DMSO reductase superfamily, catalyzes the first step of the denitrification process in bacteria. In this heterodimer, a di-heme NapB subunit is associated to the catalytic NapA subunit that binds a [4Fe-4S] cluster and a bis(molybdopterin guanine dinucleotide)...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi700928m

    authors： Dementin S,Arnoux P,Frangioni B,Grosse S,Léger C,Burlat B,Guigliarelli B,Sabaty M,Pignol D

    更新日期：2007-08-28 00:00:00

  • Interdomain Linker Determines Primarily the Structural Stability of Dystrophin and Utrophin Tandem Calponin-Homology Domains Rather than Their Actin-Binding Affinity.

    abstract：:Tandem calponin-homology (CH) domains are the most common actin-binding domains in proteins. However, structural principles underlying their function are poorly understood. These tandem domains exist in multiple conformations with varying degrees of inter-CH-domain interactions. Dystrophin and utrophin tandem CH domai...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/acs.biochem.5b00741

    authors： Bandi S,Singh SM,Mallela KM

    更新日期：2015-09-08 00:00:00

  • Homogeneity and variability in the structure of azurin molecules studied by fluorescence decay and circular polarization.

    abstract：:The fluorescence decay of apoazurin derived from Pseudomonas aeruginosa is monoexponential. By this criterion the population of molecules of apoazurin is homogeneous. The emission anisotropy factor and the absorption anisotropy factor at the red edge of the absorption band assume similar values, showing that the trypt...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00680a018

    authors： Grinvald A,Schlessinger J,Pecht I,Steinberg IZ

    更新日期：1975-05-06 00:00:00

  • The A-state of barnase.

    abstract：:The acid-induced denaturation of barnase and its mutants has been analyzed to search for partly-folded intermediates. Differential scanning calorimetry of barnase deviates from two-state behavior below pH 4.0 at low ionic strength, with the maximum discrepancy at pH 2.7. Addition of 200 mM KCl apparently restores the ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00203a015

    authors： Sanz JM,Johnson CM,Fersht AR

    更新日期：1994-09-20 00:00:00

  • Activation of isolated NADH:ubiquinone reductase I (complex I) from Escherichia coli by detergent and phospholipids. Recovery of ubiquinone reductase activity and changes in EPR signals of iron-sulfur clusters.

    abstract：:NADH:ubiquinone oxidoreductase (NDH-1 or complex I) from Escherichia coli was purified using a combination of anion exchange chromatography and centrifugation in sucrose density gradient. The dependence of enzyme activity on detergent and phospholipids was studied. Artificial hexaammineruthenium reductase activity was...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi050134v

    authors： Sinegina L,Wikström M,Verkhovsky MI,Verkhovskaya ML

    更新日期：2005-06-14 00:00:00

  • Stability effects of increasing the hydrophobicity of solvent-exposed side chains in staphylococcal nuclease.

    abstract：:A total of fifty single site surface phenylalanine substitution mutants have been made in the model protein staphylococcal nuclease. The fifty residues that were replaced with phenylalanine were chosen to give a broad sampling of solvent accessibility, secondary structure, and backbone conformations. The change in the...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi9725069

    authors： Schwehm JM,Kristyanne ES,Biggers CC,Stites WE

    更新日期：1998-05-12 00:00:00

  • Halogen Bonding: A Powerful Tool for Modulation of Peptide Conformation.

    abstract：:Halogen bonding is a weak chemical force that has so far mostly found applications in crystal engineering. Despite its potential for use in drug discovery, as a new molecular tool in the direction of molecular recognition events, it has rarely been assessed in biopolymers. Motivated by this fact, we have developed a p...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/acs.biochem.7b00429

    authors： Danelius E,Andersson H,Jarvoll P,Lood K,Gräfenstein J,Erdélyi M

    更新日期：2017-06-27 00:00:00

  • Study of the spin-spin interactions between the metal centers of Desulfovibrio gigas aldehyde oxidoreductase: identification of the reducible sites of the [2Fe-2S]1+,2+ clusters.

    abstract：:The aldehyde oxidoreductase from Desulfovibrio gigas belongs to the family of molybdenum hydroxylases. Besides a molybdenum cofactor which constitutes their active site, these enzymes contain two [2Fe-2S](2+,1+) clusters which are believed to transfer the electrons provided by the substrate to an acceptor which is eit...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi0510025

    authors： More C,Asso M,Roger G,Guigliarelli B,Caldeira J,Moura J,Bertrand P

    更新日期：2005-08-30 00:00:00

  • Pressure-induced effects on cytochrome oxidase: the aerobic steady state.

    abstract：:If cytochrome c oxidase is subjected to pressure during the aerobic steady state, large spectral changes are apparent. These seem to be associated with the inhibition of electron transport within the oxidase. The volume change for the transition is about 80 mL/mol. When the oxidase in the aerobic steady state, with po...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00414a026

    authors： Kornblatt JA,Hui Bon Hoa G,Heremans K

    更新日期：1988-07-12 00:00:00

  • Calcium ionophore increases amyloid beta peptide production by cultured cells.

    abstract：:Amyloid beta peptide (A beta) is released into the media of a variety of cells in culture during normal metabolism. The discovery of several missense mutations within or flanking the A beta region of the beta amyloid precursor protein (beta APP) in familial Alzheimer's disease provides strong evidence for a role of al...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00181a016

    authors： Querfurth HW,Selkoe DJ

    更新日期：1994-04-19 00:00:00

  • Selective binding to polynucleotides of the hybrid intercalating groove binder bis(pyrrolecarboxamide)-oxazolopyridocarbazole: a molecular modeling study.

    abstract：:In order to further characterize the binding of the hybrid molecule NetOPC [bis(pyrrolecarboxamide)-oxazolopyridocarbazole conjugate] to double-stranded DNA, we have performed a molecular modeling study to investigate the binding modes of the complexes possibly formed between NetOPC and synthetic polynucleotides poly ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00172a018

    authors： Goulaouic H,Carteau S,Subra F,Mouscadet JF,Auclair C,Sun JS

    更新日期：1994-02-15 00:00:00

  • Detection, sizing, and quantitation of polyadenylated ribonucleic acid in the nanogram-picogram range.

    abstract：:A method is described for using very high specific activity [3H]poly(deoxythymidylate) [[3H]poly(dT)] to detect, size, and quantiate subnanogram amounts of nonradioactive polyadenylated RNA. Short (approximately 100 nucleotides long) [3H]poly(dT) is hybridized to the poly(adenylate) [poly(A)] tracts in polyadenylated ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00557a022

    authors： Tracy S,Kohne DE

    更新日期：1980-08-05 00:00:00