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Weifeng Luo, Radoslav Janoštiak, Ondřej Tolde, Larisa M Ryzhova, Lenka Koudelková, Michal Dibus, Jan Brábek, Steven K Hanks, Daniel Rosel
The tyrosine kinase Src acts as a key regulator of cell motility by phosphorylating multiple protein substrates that control cytoskeletal and adhesion dynamics. In an earlier phosphotyrosine proteomics study, we identified a novel Rho-GTPase activating protein, now known as ARHGAP42, as a likely biologically relevant Src substrate. ARHGAP42 is a member of a family of RhoGAPs distinguished by tandem BAR-PH domains lying N-terminal to the GAP domain. Like other family members, ARHGAP42 acts preferentially as a GAP for RhoA...
July 15, 2017: Journal of Cell Science
Kevin Chun Chan, Lanyuan Lu, Fei Sun, Jun Fan
ACAP1 proteins were previously reported to specifically bind PIP2-containing cell membranes and form well-structured protein lattices in order to conduct membrane tubulation. We carried out molecular dynamics simulations to characterize orientation of the PH domains with respect to the BAR domains inside the protein dimer. Followed by molecular dynamics simulations, we present a comprehensive orientation analysis of PH domain under different states including unbound and bound with lipids. We have examined two binding pockets on the PH domain and present PMF profiles of the two pockets to account for their preference to PIP2 lipids...
April 20, 2017: Journal of Physical Chemistry. B
Pei-Wen Chen, Xiaoying Jian, Sarah M Heissler, Kang Le, Ruibai Luo, Lisa M Jenkins, Attila Nagy, Joel Moss, James R Sellers, Paul A Randazzo
ASAP1 regulates F-actin-based structures and functions, including focal adhesions (FAs) and circular dorsal ruffles (CDRs), cell spreading and migration. ASAP1 function requires its N-terminal BAR domain. We discovered that nonmuscle myosin 2A (NM2A) directly bound the BAR-PH tandem of ASAP1in vitro ASAP1 and NM2A co-immunoprecipitated and colocalized in cells. Knockdown of ASAP1 reduced colocalization of NM2A and F-actin in cells. Knockdown of ASAP1 or NM2A recapitulated each other's effects on FAs, cell migration, cell spreading, and CDRs...
April 1, 2016: Journal of Biological Chemistry
Decibel Elpa, Enrique Durán-Guerrero, Remedios Castro, Ramón Natera, Carmelo G Barroso
A fast, simple, and reliable analytical method for the determination of medium-level volatile thiols in wines is presented. Stir bar sorptive extraction using ethylene glycol-silicone coated stir bars has been used in combination with thermal desorption gas chromatography with mass spectrometry for the analysis of 4-mercapto-4-methylpentan-2-one, 2-furanmethanethiol, 3-mercaptohexyl acetate, and 3-mercaptohexanol in wine. Optimization of the extraction technique was performed using a two-level fractional factorial design...
July 2014: Journal of Separation Science
Marco Dias, Cédric Blanc, Nelcy Thazar-Poulot, Sabrina Ben Larbi, Pierre Cosson, François Letourneur
ACAPs and ASAPs are Arf-GTPase-activating proteins with BAR, PH, GAP and ankyrin repeat domains and are known to regulate vesicular traffic and actin cytoskeleton dynamics in mammalian cells. The amoeba Dictyostelium has only two proteins with this domain organization, instead of the six in human, enabling a more precise functional analysis. Genetic invalidation of acapA resulted in multinucleated cells with cytokinesis defects. Mutant acapA(-) cells were hardly motile and their multicellular development was significantly delayed...
February 1, 2013: Journal of Cell Science
Pei-Wen Chen, Paul A Randazzo, Carole A Parent
Arfs and Arf GTPase-activating proteins (ArfGAPs) are regulators of membrane trafficking and actin dynamics in mammalian cells. In this study, we identified a primordial Arf, ArfA, and two ArfGAPs (ACAP-A/B) containing BAR, PH, ArfGAP and Ankyrin repeat domains in the eukaryote Dictyostelium discoideum. In vitro, ArfA has similar nucleotide binding properties as mammalian Arfs and, with GTP bound, is a substrate for ACAP-A and B. We also investigated the physiological functions of ACAP-A/B by characterizing cells lacking both ACAP-A and B...
January 7, 2010: PloS One
Xiaoying Jian, Patrick Brown, Peter Schuck, James M Gruschus, Andrea Balbo, Jenny E Hinshaw, Paul A Randazzo
ASAP1 is an Arf GTPase-activating protein (GAP) that functions on membrane surfaces to catalyze the hydrolysis of GTP bound to Arf. ASAP1 contains a tandem of BAR, pleckstrin homology (PH), and Arf GAP domains and contributes to the formation of invadopodia and podosomes. The PH domain interacts with the catalytic domain influencing both the catalytic and Michaelis constants. Tandem BAR-PH domains have been found to fold into a functional unit. The results of sedimentation velocity studies were consistent with predictions from homology models in which the BAR and PH domains of ASAP1 fold together...
January 16, 2009: Journal of Biological Chemistry
Rie Ito, Migaku Kawaguchi, Norihiro Sakui, Hidehiro Honda, Noriya Okanouchi, Koichi Saito, Hiroyuki Nakazawa
A method for mercury analysis and speciation in drinking water was developed, which involved stir bar sorptive extraction (SBSE) with in situ propyl derivatization and thermal desorption (TD)-GC-MS. Ten millilitre of tap water or bottled water was used. After a stir bar, pH adjustment agent and derivatization reagent were added, SBSE was performed. Then, the stir bar was subjected to TD-GC-MS. The detection limits were 0.01 ng mL(-1) (ethylmercury; EtHg), 0.02 ng mL(-1) (methylmercury; MeHg), and 0.2 ng mL(-1) (Hg(II) and diethylmercury (DiEtHg))...
October 31, 2008: Journal of Chromatography. A
Ye-Ling Yu, Yan Lu, Xing Tang, Fu-De Cui
The purpose of this study was to prepare and evaluate the intravenous emulsion (BCOE) containing Brucea javanica oil (BJO) and Coix seed oil (CSO), which is used in anti-tumor treatment. The formulation and preparation of BCOE were systematically investigated. High-pressure homogenization, particle size distribution, zeta-potential and HPLC were carried out. The pharmacokinetics of the main component, oleic acid, and anti-tumor activity studies about the tumor growth inhibitory ratios (TGIR) and the mortality experiments were also employed to evaluate BCOE in vivo compared with BJO emulsion (BJOE) and CSO emulsion (CSOE) using S180 sarcoma-bearing mice...
April 2008: Biological & Pharmaceutical Bulletin
Heidi J Chial, Ruping Wu, Carolyn V Ustach, Linda C McPhail, William C Mobley, Yong Q Chen
Human adaptor protein, phosphotyrosine interaction, PH domain and leucine zipper containing 1 (APPL1) and adaptor protein, phosphotyrosine interaction, PH domain and leucine zipper containing 2 (APPL2) are homologous effectors of the small guanosine triphosphatase RAB5 that interact with a diverse set of receptors and signaling proteins and are proposed to function in endosome-mediated signaling. Herein, we investigated the membrane-targeting properties of the APPL1 and APPL2 Bin/Amphiphysin/Rvs (BAR), pleckstrin homology (PH) and phosphotyrosine binding (PTB) domains...
February 2008: Traffic
Guangyu Zhu, Jia Chen, Jay Liu, Joseph S Brunzelle, Bo Huang, Nancy Wakeham, Simon Terzyan, Xuemei Li, Zihe Rao, Guangpu Li, Xuejun C Zhang
APPL1 is an effector of the small GTPase Rab5. Together, they mediate a signal transduction pathway initiated by ligand binding to cell surface receptors. Interaction with Rab5 is confined to the amino (N)-terminal region of APPL1. We report the crystal structures of human APPL1 N-terminal BAR-PH domain motif. The BAR and PH domains, together with a novel linker helix, form an integrated, crescent-shaped, symmetrical dimer. This BAR-PH interaction is likely conserved in the class of BAR-PH containing proteins...
July 25, 2007: EMBO Journal
Jiang Li, Xuming Mao, Lily Q Dong, Feng Liu, Liang Tong
APPL1 interacts with adiponectin receptors and other important signaling molecules. It contains a BAR and a PH domain near its N terminus, and the two domains may function as a unit (BAR-PH domain). We report here the crystal structures of the BAR-PH and PTB domains of human APPL1. The structures reveal novel features for BAR domain dimerization and for the interactions between the BAR and PH domains. The BAR domain dimer of APPL1 contains two four-helical bundles, whereas other BAR domain dimers have only three helices in each bundle...
May 2007: Structure
Masuko Katoh, Masaru Katoh
ARHGAP family genes encode Rho/Rac/Cdc42-like GTPase activating proteins with RhoGAP domain. Here, we characterized human ARHGAP10 gene by using bioinformatics. Complete coding sequence of ARHGAP10 isoform A was determined by assembling nucleotide position 1-725 of FLJ41791 cDNA (AK123785.1) and 5'-truncated IMAGE4310652 cDNA (BC011920.2). Nucleotide position 240-2600 of ARHGAP10 isoform A was identical to GRAF2 cDNA (AB050785.1). Complete coding sequence of ARHGAP10 isoform B was derived from FLJ41791 cDNA...
October 2004: International Journal of Oncology
R A Alberty
The thermodynamics of six reactions of nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate is discussed both from the viewpoint of the chemical equations and the biochemical equations for these reactions. Tables of the standard enthalpies of formation and standard Gibbs energies of formation of species are presented and are used to calculate standard enthalpies of reaction, standard Gibbs energies of reaction, and equilibrium constants K of chemical reactions at 25 degrees C, 1 bar, and ionic strengths of 0, 0...
November 15, 1993: Archives of Biochemistry and Biophysics
R A Alberty
The thermodynamics of the nitrogenase reactions are discussed in terms of chemical equations and biochemical equations. Chemical equations balance all elements and electric charge. Biochemical equations represent changes at specified pH and specified free concentrations of metal ions that are bound by reactants, but they do not balance hydrogen or metal ions that have specified free concentrations. At a specified pH, it takes three separate biochemical equations to represent the changes catalyzed by nitrogenase...
March 11, 1994: Journal of Biological Chemistry
H D Sgan-Cohen, E Newbrun, R Huber, G Tenebaum, M N Sela
In this study we investigated the effect of previous diet on the response of plaque pH to three test foods. The study population consisted of 11 dental students. Plaque pH was measured, by means of the touch electrode method, on the first two upper molars. The study was conducted at two sessions, one week apart. Subjects followed a 48-hour high-sugar diet before the first test session and a 48-hour low-sugar diet before the second test session. During both 48-hour periods, students refrained from all oral hygiene practices and fluoride utilization...
November 1988: Journal of Dental Research
R A Alberty, R N Goldberg
The criterion for chemical equilibrium at specified temperature, pressure, pH, concentration of free magnesium ion, and ionic strength is the transformed Gibbs energy, which can be calculated from the Gibbs energy. The apparent equilibrium constant (written in terms of the total concentrations of reactants like adenosine 5'-triphosphate, rather than in terms of species) yields the standard transformed Gibbs energy of reaction, and the effect of temperature on the apparent equilibrium constant at specified pressure, pH, concentration of free magnesium ion, and ionic strength yields the standard transformed enthalpy of reaction...
November 3, 1992: Biochemistry
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