Mutational analysis reveals a single binding interface between RhoA and its effector, PRK1.


:Protein kinase C-related kinases (PRKs) are serine/threonine kinases that are members of the protein kinase C superfamily and can be activated by binding to members of the Rho family of small G proteins via a Rho binding motif known as an HR1 domain. The PRKs contain three tandem HR1 domains at their N-termini. The structure of the HR1a domain from PRK1 in complex with RhoA [Maesaki, R., et al. (1999) Mol. Cell 4, 793-803] identified two potential contact interfaces between the G protein and the HR1a domain. In this work, we have used an alanine scanning mutagenesis approach to identify whether both contact sites are used when the two proteins interact in solution and also whether HR1b, the second HR1 domain from PRK1, plays a role in binding to RhoA. The mutagenesis identified just one contact site as being relevant for binding of RhoA and HR1a in solution, and the HR1b domain was found not to contribute to RhoA binding. The folded state and thermal stability of the HR1a and HR1b domains were also investigated. HR1b was found to be more thermally stable than HR1a, and it is hypothesized that the differences in the biophysical properties of these two domains govern their interaction with small G proteins.






Hutchinson CL,Lowe PN,McLaughlin SH,Mott HR,Owen D




Has Abstract


2011-04-12 00:00:00












  • Overexpression and divalent metal binding properties of the methionyl aminopeptidase from Pyrococcus furiosus.

    abstract::The gene encoding for the methionyl aminopeptidase from the hyperthermophilic archaeon Pyrococcus furiosus (PfMetAP-II; EC has been inserted into a pET 27b(+) vector and overexpressed in Escherichia coli. The new expression system resulted in a 5-fold increase in purified enzyme obtained from a 5 L fermento...


    pub_type: 杂志文章


    authors: Meng L,Ruebush S,D'souza VM,Copik AJ,Tsunasawa S,Holz RC

    更新日期:2002-06-11 00:00:00

  • Substantial Decrease in Plasmalogen in the Heart Associated with Tafazzin Deficiency.

    abstract::Tafazzin is the mitochondrial enzyme that catalyzes transacylation between a phospholipid and a lysophospholipid in remodeling. Mutations in tafazzin cause Barth syndrome, a potentially life-threatening disease with the major symptom being cardiomyopathy. In the tafazzin-deficient heart, cardiolipin (CL) acyl chains b...


    pub_type: 杂志文章


    authors: Kimura T,Kimura AK,Ren M,Berno B,Xu Y,Schlame M,Epand RM

    更新日期:2018-04-10 00:00:00

  • Heme-CO religation in photolyzed hemoglobin: a time-resolved Raman study of the Fe-CO stretching mode.

    abstract::Time-resolved resonance Raman spectroscopy has been employed to monitor geminate heme-CO rebinding in photolyzed HbCO. The excitation frequency was tuned to enhance the scattering from rebound heme sites 20-500 ns subsequent to CO photolysis. The behavior of vFe-C during ligand rebinding has important ramifications co...


    pub_type: 杂志文章


    authors: Schneebeck MC,Vigil LE,Friedman JM,Chavez MD,Ondrias MR

    更新日期:1993-02-09 00:00:00

  • The androgen receptor T877A mutant recruits LXXLL and FXXLF peptides differently than wild-type androgen receptor in a time-resolved fluorescence resonance energy transfer assay.

    abstract::The interactions of the ligand binding domain (LBD) of androgen receptor (AR) and the AR T877A mutant, found in prostate cancer, with peptides from coactivator and corepressor proteins or random phage display peptides were investigated using in vitro time-resolved fluorescence resonance energy transfer (TR-FRET). Inte...


    pub_type: 杂志文章


    authors: Ozers MS,Marks BD,Gowda K,Kupcho KR,Ervin KM,De Rosier T,Qadir N,Eliason HC,Riddle SM,Shekhani MS

    更新日期:2007-01-23 00:00:00

  • Free energy rhythms in Saccharomyces cerevisiae: a dynamic perspective with implications for ribosomal biogenesis.

    abstract::To describe the time course of cellular systems, we integrate ideas from thermodynamics and information theory to discuss the work needed to change the state of the cell. The biological example analyzed is experimental microarray transcription level oscillations of yeast in the different phases as characterized by oxy...


    pub_type: 杂志文章


    authors: Gross A,Li CM,Remacle F,Levine RD

    更新日期:2013-03-05 00:00:00

  • Generation and characterization of two transgenic mouse lines expressing human ApoE2 in neurons and glial cells.

    abstract::Apolipoprotein E (apoE) isoforms are key determinants of susceptibility to late-onset Alzheimer's disease (AD). The epsilon 4 and epsilon 2 alleles have been associated with increased and decreased risk for AD, respectively. We have generated and characterized transgenic mice in which the human apoE2 gene is expressed...


    pub_type: 杂志文章


    authors: Georgopoulos S,McKee A,Kan HY,Zannis VI

    更新日期:2002-07-30 00:00:00

  • Importance of central alpha-helices of human apolipoprotein A-I in the maturation of high-density lipoproteins.

    abstract::We have studied the role of amphipathic alpha-helices in the ability of apoA-I to promote cholesterol efflux from human skin fibroblasts and activate lecithin:cholesterol acyltransferase (LCAT). Three apoA-I mutants were designed, each by deletion of a pair of predicted adjacent central alpha-helices [Delta(100-143), ...


    pub_type: 杂志文章


    authors: Frank PG,N'Guyen D,Franklin V,Neville T,Desforges M,Rassart E,Sparks DL,Marcel YL

    更新日期:1998-09-29 00:00:00

  • Structure of a specialized acyl carrier protein essential for lipid A biosynthesis with very long-chain fatty acids in open and closed conformations.

    abstract::The solution nuclear magnetic resonance (NMR) structures and backbone (15)N dynamics of the specialized acyl carrier protein (ACP), RpAcpXL, from Rhodopseudomonas palustris, in both the apo form and holo form modified by covalent attachment of 4'-phosphopantetheine at S37, are virtually identical, monomeric, and corre...


    pub_type: 杂志文章


    authors: Ramelot TA,Rossi P,Forouhar F,Lee HW,Yang Y,Ni S,Unser S,Lew S,Seetharaman J,Xiao R,Acton TB,Everett JK,Prestegard JH,Hunt JF,Montelione GT,Kennedy MA

    更新日期:2012-09-18 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...


    pub_type: 杂志文章


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

    更新日期:1988-07-12 00:00:00

  • Library Selection with a Randomized Repertoire of (βα)8-Barrel Enzymes Results in Unexpected Induction of Gene Expression.

    abstract::The potential of the frequently encountered (βα)8-barrel fold to acquire new functions was tested by an approach combining random mutagenesis and selection in vivo. For this purpose, the genes encoding 52 different phosphate-binding (βα)8-barrel proteins were subjected to error-prone PCR and cloned into an expression ...


    pub_type: 杂志文章


    authors: Rohweder B,Lehmann G,Eichner N,Polen T,Rajendran C,Ruperti F,Linde M,Treiber T,Jung O,Dettmer K,Meister G,Bott M,Gronwald W,Sterner R

    更新日期:2019-10-15 00:00:00

  • Electron spin echo envelope modulation study of oxygenated iron-cobalt hybrid hemoglobins reveals molecular features analogous to those of the oxy ferrous protein.

    abstract::Two oxygenated iron-cobalt hybrid hemoglobins (Hbs), (alpha Co-O2 beta Fe-O2)2 and (alpha Fe-O2 beta Co-O2)2, were studied by electron spin echo envelope modulation (ESEEM) spectroscopy in order to measure (i) electron-nuclear hyperfine and nuclear quadrupole coupling to the N epsilon of the proximal histidyl imidazol...


    pub_type: 杂志文章


    authors: Lee HC,Peisach J,Tsuneshige A,Yonetani T

    更新日期:1995-05-23 00:00:00

  • Glutathione reductase turned into trypanothione reductase: structural analysis of an engineered change in substrate specificity.

    abstract::Trypanosoma and Leishmania, pathogens responsible for diseases such as African sleeping sickness, Chagas' heart disease, or Oriental sore, are two of the very few genera that do not use the ubiquitous glutathione/glutathione reductase system to keep a stable cellular redox balance. Instead, they rely on trypanothione ...


    pub_type: 杂志文章


    authors: Stoll VS,Simpson SJ,Krauth-Siegel RL,Walsh CT,Pai EF

    更新日期:1997-05-27 00:00:00

  • Mechanism of autophosphorylation of mycobacterial PknB explored by molecular dynamics simulations.

    abstract::Mycobacterial Ser/Thr kinase, PknB, is essential for the growth of the pathogen. Unphosphorylated PknB is catalytically inactive, and its activation requires autophosphorylation of Thr residues on the activation loop. Autophosphorylation can in principle take place via two distinct mechanisms. Intermolecular trans aut...


    pub_type: 杂志文章


    authors: Damle NP,Mohanty D

    更新日期:2014-07-22 00:00:00

  • Amino acid and sequence analysis of the cytochrome and flavoprotein subunits of p-cresol methylhydroxylase.

    abstract::The flavocytochrome p-cresol methylhydroxylase from Pseudomonas putida has been reported to have a Mr of 114,000 and to consist of two subunits, a flavoprotein and a cytochrome c, each with a Mr of 58,000. Recent X-ray crystallographic data from our laboratories [Shamala, N., Lim, L. W., Mathews, F. S., McIntire, W., ...


    pub_type: 杂志文章


    authors: McIntire W,Singer TP,Smith AJ,Mathews FS

    更新日期:1986-10-07 00:00:00

  • Overexpression, purification, and mechanistic study of UDP-N-acetylenolpyruvylglucosamine reductase.

    abstract::The recently isolated Escherichia coli murB gene (Pucci et al., 1992) has been cloned into an expression vector and the encoded UDP-N-acetylenolpyruvylglucosamine reductase (EC was overproduced to about 10% of soluble cell protein. The encoded 38-kDa protein has been purified to near homogeneity. It was fou...


    pub_type: 杂志文章


    authors: Benson TE,Marquardt JL,Marquardt AC,Etzkorn FA,Walsh CT

    更新日期:1993-03-02 00:00:00

  • Mutation of Tyr138 disrupts the structural coupling between the opposing domains in vertebrate calmodulin.

    abstract::We have used circular dichroism and frequency-domain fluorescence spectroscopy to determine how the site-specific substitution of Tyr138 with either Phe138 or Gln138 affects the structural coupling between the opposing domains of calmodulin (CaM). A double mutant was constructed involving conservative substitution of ...


    pub_type: 杂志文章


    authors: Sun H,Yin D,Coffeen LA,Shea MA,Squier TC

    更新日期:2001-08-14 00:00:00

  • Anaerobic Heme Degradation: ChuY Is an Anaerobilin Reductase That Exhibits Kinetic Cooperativity.

    abstract::Heme catabolism is an important biochemical process that many bacterial pathogens utilize to acquire iron. However, tetrapyrrole catabolites can be reactive and often require further processing for transport out of the cell or conversion to another useful cofactor. In previous work, we presented in vitro evidence of a...


    pub_type: 杂志文章


    authors: LaMattina JW,Delrossi M,Uy KG,Keul ND,Nix DB,Neelam AR,Lanzilotta WN

    更新日期:2017-02-14 00:00:00

  • Isolation and structural determination of novel sulfated hexasaccharides from squid cartilage chondroitin sulfate E that exhibits neuroregulatory activities.

    abstract::Squid cartilage chondroitin sulfate E (CS-E) exhibits various biological activities, including anticoagulant activities, lymphoid regulatory activities, and neuroregulatory activities [Ueoka, C., Kaneda, N., Okazaki, I., Nadanaka, S., Muramatsu, T., and Sugahara, K. (2000) J. Biol. Chem. 275, 37407-37413]. These activ...


    pub_type: 杂志文章


    authors: Kinoshita A,Yamada S,Haslam SM,Morris HR,Dell A,Sugahara K

    更新日期:2001-10-23 00:00:00

  • Dynamics of proton diffusion within the hydration layer of phospholipid membrane.

    abstract::The diffusion of protons at the immediate vicinity of (less than 10 A from) a phospholipid membrane is studied by the application of the laser-induced proton pulse. A light-sensitive proton emitter (8-hydroxypyrene-1,3,6-trisulfonate) was trapped exclusively in the hydration layers of multilamellar vesicles made of eg...


    pub_type: 杂志文章


    authors: Gutman M,Nachliel E,Moshiach S

    更新日期:1989-04-04 00:00:00

  • Thermodynamic and kinetic examination of protein stabilization by glycerol.

    abstract::The effect of concentrated glycerol on the thermal transitions of chymotrypsinogen and ribonuclease has been examined by differential spectrophotometry at 293 and 287 mm, respectively. It was found that for both proteins addition of glycerol raises the transition temperature, the increase in Tm being greater for ribon...


    pub_type: 杂志文章


    authors: Gekko K,Timasheff SN

    更新日期:1981-08-04 00:00:00

  • Mechanism-based inactivation of cytochrome P450 2B4 by aldehydes: relationship to aldehyde deformylation via a peroxyhemiacetal intermediate.

    abstract::The inactivation of cytochrome P450 2B4 by aldehydes in a reconstituted enzyme system requires molecular oxygen and NADPH and is not prevented by the addition of catalase, superoxide dismutase, epoxide hydrolase, glutathione, or ascorbic acid. A strong correlation between loss of enzymatic activity and bleaching of th...


    pub_type: 杂志文章


    authors: Raner GM,Chiang EW,Vaz AD,Coon MJ

    更新日期:1997-04-22 00:00:00

  • How fumarase recycles after the malate --> fumarate reaction. Insights into the reaction mechanism.

    abstract::Recycling of yeast fumarase to permit repetition of its reaction chemistry requires two proton transfers and two conformational changes, in pathways that are different in detail but thematically similar in the two directions. In the malate --> fumarate direction, simple anions such as acetate accelerate the fumarate-o...


    pub_type: 杂志文章


    authors: Rose IA

    更新日期:1998-12-22 00:00:00

  • Effects of metal ligation and oxygen on the reversibility of the thermal denaturation of Pseudomonas aeruginosa azurin.

    abstract::Thermodynamic equilibrium transition models in DSC are only applicable to reversible processes, but reversibility of the thermal transitions of proteins is comparatively rare because of intermolecular aggregation of denatured proteins and the degradation that occurs at high temperatures. The cupredoxin azurin from Pse...


    pub_type: 杂志文章


    authors: Sandberg A,Leckner J,Shi Y,Schwarz FP,Karlsson BG

    更新日期:2002-01-22 00:00:00

  • The heparin binding domain of S-protein/vitronectin binds to complement components C7, C8, and C9 and perforin from cytolytic T-cells and inhibits their lytic activities.

    abstract::S-Protein/vitronectin is a serum glycoprotein that inhibits the lytic activity of the membrane attack complex of complement, i.e., of the complex including the proteins C5b, C6, C7, C8, and C9n. We show that intact S-protein/vitronectin or its cyanogen bromide generated fragments also inhibit the hemolysis mediated by...


    pub_type: 杂志文章


    authors: Tschopp J,Masson D,Schäfer S,Peitsch M,Preissner KT

    更新日期:1988-05-31 00:00:00

  • tRNA and the guanosinetriphosphatase activity of elongation factor Tu.

    abstract::Different sites of the tRNA molecule influence the activity of the elongation factor Tu (EF-Tu) center for GTP hydrolysis [Parlato, G., Pizzano, R., Picone, D., Guesnet, J., Fasano, O., & Parmeggiani, A. (1983) J. Biol. Chem. 258, 995-1000]. Continuing these studies, we have investigated some aspects of (a) the effect...


    pub_type: 杂志文章


    authors: Swart GW,Parmeggiani A

    更新日期:1989-01-10 00:00:00

  • Differential susceptibility of transferrin glycoforms to chymotrypsin: a proteomics approach to the detection of carbohydrate-deficient transferrin.

    abstract::Transferrin exhibits heterogeneity in glycosylation characteristic of pathological changes in alcohol abuse and congenital disorders in glycosylation. This study investigated an alternative approach in the detection of carbohydrate-deficient transferrin based on the premise that glycosylation may afford some degree of...


    pub_type: 杂志文章


    authors: Valmu L,Kalkkinen N,Husa A,Rye PD

    更新日期:2005-12-13 00:00:00

  • Investigation into the catalytic role for the tryptophan residues within domain III of Pseudomonas aeruginosa exotoxin A.

    abstract::The role of the tryptophan residues in the substrate-binding and catalytic mechanism of an enzymatically active C-terminal fragment of Pseudomonas aeruginosa exotoxin A was studied by individually or jointly replacing these residues with phenylalanine. Substitution of W-466 decreased the ADP-ribosyltransferase and NAD...


    pub_type: 杂志文章


    authors: Beattie BK,Prentice GA,Merrill AR

    更新日期:1996-12-03 00:00:00

  • Aminoacyl thioester chemistry of class II aminoacyl-tRNA synthetases.

    abstract::Lysyl-tRNA synthetase, a class II enzyme, edits homocysteine by converting it into homocysteine thiolactone. In a similar reaction, the enzyme converts homoserine into homoserine lactone. Other class II enzymes, aspartyl-tRNA synthetase and seryl-tRNA synthetase, do not edit any of the amino acids tested. However, all...


    pub_type: 杂志文章


    authors: Jakubowski H

    更新日期:1997-09-16 00:00:00

  • Molecular model of equine infectious anemia virus proteinase and kinetic measurements for peptide substrates with single amino acid substitutions.

    abstract::A molecular model has been built of the equine infectious anemia virus (EIAV) proteinase on the basis of the crystal structures of the related Rous sarcoma virus (RSV) and human immunodeficiency virus (HIV) proteinases. The 104 residue long EIAV proteinase has 30 identical and 11 similar amino acids compared to those ...


    pub_type: 杂志文章


    authors: Weber IT,Tözsér J,Wu J,Friedman D,Oroszlan S

    更新日期:1993-04-06 00:00:00

  • Hypoxia Inducible Factors Modulate Mitochondrial Oxygen Consumption and Transcriptional Regulation of Nuclear-Encoded Electron Transport Chain Genes.

    abstract::Hypoxia inducible factor-1 (HIF1) is a stress-responsive nuclear transcription factor that is activated with a decrease in oxygen availability. HIF1 regulates the expression of genes involved in a cell's adaptation to hypoxic stress, including those with mitochondrial specific function. To gain a more comprehensive un...


    pub_type: 杂志文章


    authors: Hwang HJ,Lynn SG,Vengellur A,Saini Y,Grier EA,Ferguson-Miller SM,LaPres JJ

    更新日期:2015-06-23 00:00:00