Details of mannitol transport in Escherichia coli elucidated by site-specific mutagenesis and complementation of phosphorylation site mutants of the phosphoenolpyruvate-dependent mannitol-specific phosphotransferase system.


:The mannitol transport protein (EIImtl) carries out translocation with concomitant phosphorylation of mannitol from the periplasm to the cytoplasm, at the expense of phosphoenolpyruvate (PEP). The phosphoryl group which is needed for this group translocation is sequentially transferred from PEP via two phosphorylation sites, located exclusively on the C-terminal cytoplasmic domain, to mannitol. Oligonucleotide-directed mutagenesis was used to investigate the precise role of these sites in phosphoryl group transfer, by producing specific amino acid substitutions. The first phosphorylation site, His-554 (P1), was replaced by Ala, which renders the EII-H554A completely inactive in PEP-dependent mannitol phosphorylation, but not in mannitol/mannitol 1-phosphate exchange. The P2 site mutant, EII-C384S, was inactive both in the mannitol phosphorylation reaction and in the exchange reaction, due to replacement of the essential Cys-384 by Ser. Although EII-H554A and EII-C384S were both catalytically inactive in the PEP-dependent phosphorylation, EII-C384S was able to restore up to 55% of the wild-type mannitol phosphorylation activity with the EII-H554A mutant, indicating a direct phosphotransfer between two subunits. These phosphorylation data together with the data obtained from mannitol/mannitol phosphate exchange kinetics, after mixing EII-H554A and EII-C384S, indicated the formation of functionally stable heterodimers, which consist of an EII-H554A and an EII-C384S monomer.






van Weeghel RP,van der Hoek YY,Pas HH,Elferink M,Keck W,Robillard GT




Has Abstract


1991-02-19 00:00:00












  • Solution studies on the structure of bent DNA in the cAMP receptor protein-lac DNA complex.

    abstract::Cyclic AMP receptor protein is involved in the regulation of more than 20 genes. A step in the mechanism of activation of transcription is to induce a significant bending of the DNA upon complex formation between specific DNA and the protein. The induced DNA bending and a structure of the protein-DNA complex were stud...


    pub_type: 杂志文章


    authors: Heyduk T,Lee JC

    更新日期:1992-06-09 00:00:00

  • The structure of a soluble chemoreceptor suggests a mechanism for propagating conformational signals.

    abstract::Transmembrane chemoreceptors, also known as methyl-accepting chemotaxis proteins (MCPs), translate extracellular signals into intracellular responses in the bacterial chemotaxis system. MCP ligand binding domains control the activity of the CheA kinase, situated approximately 200 A away, across the cytoplasmic membran...


    pub_type: 杂志文章


    authors: Pollard AM,Bilwes AM,Crane BR

    更新日期:2009-03-10 00:00:00

  • Specific proteolysis of human plasminogen by a 24 kDa endopeptidase from a novel Chryseobacterium Sp.

    abstract::A novel single polypeptide endopeptidase of 24 kDa (24k-endopeptidase) was purified with a yield of 300-400 microg/L from conditioned medium of a bacterial strain which was identified as a new species in the genus Chryseobacterium Sp. on the basis of its 16S rDNA sequence and DNA:DNA hybridizations. The NH(2)-terminal...


    pub_type: 杂志文章


    authors: Lijnen HR,Van Hoef B,Ugwu F,Collen D,Roelants I

    更新日期:2000-01-18 00:00:00

  • The BARD1 C-terminal domain structure and interactions with polyadenylation factor CstF-50.

    abstract::The BARD1 N-terminal RING domain binds BRCA1 while the BARD1 C-terminal ankyrin and tandem BRCT repeat domains bind CstF-50 to modulate mRNA processing and RNAP II stability in response to DNA damage. Here we characterize the BARD1 structural biochemistry responsible for CstF-50 binding. The crystal structure of the B...


    pub_type: 杂志文章


    authors: Edwards RA,Lee MS,Tsutakawa SE,Williams RS,Nazeer I,Kleiman FE,Tainer JA,Glover JN

    更新日期:2008-11-04 00:00:00

  • Cooperative binding of polyamines induces the Escherichia coli single-strand binding protein-DNA binding mode transitions.

    abstract::The Escherichia coli single-strand binding (SSB) protein is an essential protein involved in DNA replication, recombination, and repair processes. The tetrameric protein binds to ss nucleic acids in a number of different binding modes in vitro. These modes differ in the number of nucleotides occluded per SSB tetramer ...


    pub_type: 杂志文章


    authors: Wei TF,Bujalowski W,Lohman TM

    更新日期:1992-07-07 00:00:00

  • Relevance of partially structured states in the non-classical secretion of acidic fibroblast growth factor.

    abstract::Acidic fibroblast growth factor (aFGF) is a signal peptide-less protein that is secreted into the extracellular compartment as part of a multiprotein release complex, consisting of aFGF, S100A13 (a calcium binding protein), and a 40 kDa (p40) form of synaptotagmin (Syt1), a protein that participates in the docking of ...


    pub_type: 杂志文章


    authors: Rajalingam D,Graziani I,Prudovsky I,Yu C,Kumar TK

    更新日期:2007-08-14 00:00:00

  • Differential scanning calorimetry study of local anesthetic effects on F1ATPase and submitochondrial particles.

    abstract::The differential scanning calorimetry trace of F1ATPase, prepared from beef heart submitochondrial particles, has a single sharp endothermic transition at 80.5 +/- 1.0 degrees C and a half-height peak width of 2.0 +/- 0.2 degrees. The transition enthalpy is 19 +/- 2 cal/g of protein. Submitochondrial particles (SMP) h...


    pub_type: 杂志文章


    authors: Kresheck GC,Adade AB,Vanderkooi G

    更新日期:1985-03-26 00:00:00

  • Regulation of plasminogen activation by components of the extracellular matrix.

    abstract::The kinetics of activation of Glu-plasminogen (Glu-Pg) and Lys77-Pg by two-chain recombinant tissue plasminogen activator (t-PA) were determined in the presence of isolated protein components of the extracellular matrix (ECM) and compared to activation in the presence of fibrinogen and fibrinogen fragments and in the ...


    pub_type: 杂志文章


    authors: Stack S,Gonzalez-Gronow M,Pizzo SV

    更新日期:1990-05-22 00:00:00

  • Hidden Conformational States and Strange Temperature Optima in Enzyme Catalysis.

    abstract::The existence of temperature optima in enzyme catalysis that occur before protein melting sets in can be described by different types of kinetic models. Such optima cause distinctly curved Arrhenius plots and have, for example, been observed in several cold-adapted enzymes from psychrophilic species. The two main expl...


    pub_type: 杂志文章


    authors: Åqvist J,Sočan J,Purg M

    更新日期:2020-10-13 00:00:00

  • 23,24,25-Trihydroxyvitamin D3, 24,25,26-trihydroxyvitamin D3, 24-keto-25-hydroxyvitamin D3, and 23-dehydro-25-hydroxyvitamin D3: new in vivo metabolites of vitamin D3.

    abstract::Four new in vivo metabolites of vitamin D3 were isolated from the blood plasma of chicks given large doses of vitamin D3. The metabolites were isolated by methanol-chloroform extraction and a series of chromatographic procedures. By use of mass spectrometry, ultraviolet absorption spectrophotometry, and specific chemi...


    pub_type: 杂志文章


    authors: Wichmann JK,Schnoes HK,DeLuca HF

    更新日期:1981-12-22 00:00:00

  • Raman difference spectroscopic studies of the myosin S1.MgADP.vanadate complex.

    abstract::The Raman spectra of the nonbridging V--O bonds in the myosin S1.MgADP.Vi complex, often believed to be a transition-state analogue for the phosphotransfer reaction catalyzed by myosin, and in a vanadate solution model compound have been obtained using Raman difference spectroscopic techniques. A symmetric/asymmetric ...


    pub_type: 杂志文章


    authors: Deng H,Wang J,Callender RH,Grammer JC,Yount RG

    更新日期:1998-08-04 00:00:00

  • The selectivity for K+ versus Na+ in DNA quadruplexes is dominated by relative free energies of hydration: a thermodynamic analysis by 1H NMR.

    abstract::We have studied the competition between Na+ and K+ for coordination by G quartets using the oligonucleotide d(G3T4G3) as a model system. d(G3T4G3) forms a dimeric foldback structure containing three G quartets in the presence of either NaCl or KCl. Proton chemical shifts, which are particular to the species of coordin...


    pub_type: 杂志文章


    authors: Hud NV,Smith FW,Anet FA,Feigon J

    更新日期:1996-12-03 00:00:00

  • New clevable photoreactive heterobifunctional cross-linking reagents for studying membrane organization.

    abstract::The synthesis is described of four new cleavable, photosensitive, heterobifunctional cross-linking reagents for use in examining membrane organization: 4'-azidoazobenzene-4oxysuccinimide ester (1), N-[4-(p-azidophenylazo)benzoyl]-3-aminopropyl-N'-oxysuccinimide ester (2), N-[4-(p-azidophenylazo)benzoyl]-6-aminohexyl-N...


    pub_type: 杂志文章


    authors: Jaffe CL,Lis H,Sharon N

    更新日期:1980-09-16 00:00:00

  • Mutational analysis of the role of the N terminus of actin in actomyosin interactions. Comparison with other mutant actins and implications for the cross-bridge cycle.

    abstract::Yeast actin mutants with acidic residues at the N terminus either neutralized (DNEQ) or deleted (delta-DSE) were used to assess the role of N-terminal acidic residues in the interactions of actin with myosin in the contractile cycle. Cosedimentation experiments revealed an approximately 3-fold decrease in the binding ...


    pub_type: 杂志文章


    authors: Miller CJ,Wong WW,Bobkova E,Rubenstein PA,Reisler E

    更新日期:1996-12-24 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...


    pub_type: 杂志文章


    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

  • Pump-1 cDNA codes for a protein with characteristics similar to those of classical collagenase family members.

    abstract::Pump-1 cDNA has recently been isolated by screening a human tumor cDNA library with a transin (rat stromelysin) probe under low-stringency hybridization conditions. The cDNA codes for a potential protein with significant sequence similarity to the metalloproteinases collagenase and stromelysin, but which lacks the hem...


    pub_type: 杂志文章


    authors: Quantin B,Murphy G,Breathnach R

    更新日期:1989-06-27 00:00:00

  • Structure and vectorial properties of proteoliposomes containing cytochrome oxidase in the submitochondrial orientation.

    abstract::Cytochrome oxidase proteoliposomes were prepared from bovine heart oxidase. Size distributions determined by quasi-elastic light scattering (QELS) showed that there was a small population of large vesicles (120-200-nm diameter) and a large population of small vesicles (50-100-nm diameter). Trapping cytochrome c inside...


    pub_type: 杂志文章


    authors: Cooper CE,Nicholls P

    更新日期:1990-04-24 00:00:00

  • Ionic and nucleotide requirements for microtubule polymerization in vitro.

    abstract::The ionic and nucleotide requirements for the in vitro polymerization of microtubules from purified brain tubulin have been characterized by viscometry. Protein was purified by successive cycles of a temperature dependent assembly-diassembly scheme. Maximal polymerization occurred at a concentration of 0.1 M Pipes (pi...


    pub_type: 杂志文章


    authors: Olmsted JB,Borisy GG

    更新日期:1975-07-01 00:00:00

  • Time-resolved fluorescence investigations of the interaction of the voltage-sensitive probe RH421 with lipid membranes and proteins.

    abstract::Fluorescence lifetimes and fluorescence anisotropy decays of the voltage-sensitive styryl-pyridinium dye RH421 have been measured in the presence of dimyristoylphosphatidylcholine vesicles, the water soluble enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (rubisco), and Na+,K(+)-ATPase-containing membrane fragm...


    pub_type: 杂志文章


    authors: Visser NV,van Hoek A,Visser AJ,Frank J,Apell HJ,Clarke RJ

    更新日期:1995-09-19 00:00:00

  • Two-dimensional NMR characterization of the deoxymyoglobin heme pocket.

    abstract::Traditionally, assigning the heme protein resonances has relied heavily on the comparison of spectra arising from protein reconstituted with specifically deuterated hemes and the native form. Such an approach can identify tentatively the broad, overlapping signals in the Fe(II) high-spin heme protein spectra. Although...


    pub_type: 杂志文章


    authors: Busse SC,Jue T

    更新日期:1994-09-13 00:00:00

  • Interaction of carbohydrate binding sites on concanavalin A-agarose with receptors on adipocytes studied by buoyant density method.

    abstract::The interaction of concanavalin A (Con A) with isolated adipocytes was studied using Con A-Sepharose beads in the affinity binding buoyant density method previously used to study insulin receptors. Free Con A-Sepharose beads could be separated from the bound beads (cell-bead complexes) by sedimentation of the high den...


    pub_type: 杂志文章


    authors: Katzen HM,Soderman DD

    更新日期:1975-06-03 00:00:00

  • Function of arginine-234 and aspartic acid-271 in domain closure, cooperativity, and catalysis in Escherichia coli aspartate transcarbamylase.

    abstract::Two mutant versions of Escherichia coli aspartate transcarbamylase were created by site-specific mutagenesis. Arg-234 of the 240s loop was replaced by serine in order to help deduce the function of the interactions that normally occur between Arg-234 and both Glu-50 and Gln-231 in the R state of the enzyme. The other ...


    pub_type: 杂志文章


    authors: Middleton SA,Kantrowitz ER

    更新日期:1988-11-15 00:00:00

  • Magnetic circular dichroism studies on the mononuclear ferrous active site of phthalate dioxygenase from Pseudomonas cepacia show a change of ligation state on substrate binding.

    abstract::Phthalate dioxygenase from Pseudomonas cepacia contains a mononuclear ferrous center that is strictly required for catalytic oxygen activation. The spectroscopic characterization of this iron site and its ligand interactions has been complicated in the past by interference from a Rieske-type binuclear (2Fe-2S) cluster...


    pub_type: 杂志文章


    authors: Gassner GT,Ballou DP,Landrum GA,Whittaker JW

    更新日期:1993-05-11 00:00:00

  • Secretion of 35SO4-labeled proteins from isolated rat hepatocytes.

    abstract::Sulfation is a Golgi-specific modification of secretory proteins. We have characterized the proteins that are labeled with 35SO4 in cultures of rat hepatocytes and studied their transport to the medium. Analysis by polyacrylamide gel electrophoresis showed that of the five most heavily labeled proteins, four had well-...


    pub_type: 杂志文章


    authors: von Würtemberg MM,Fries E

    更新日期:1989-05-02 00:00:00

  • Fluorescence study of brevin, the Mr 92 000 actin-capping and -fragmenting protein isolated from serum. Effect of Ca2+ on protein conformation.

    abstract::The fluorescence characteristics of brevin and the effects of Ca2+ on the protein conformation were fully investigated. Brevin contains 18 tryptophans and 27 tyrosines. Analysis of the fluorescence spectra and the accessibility to quenching molecules indicate that the emitting tryptophans are located in a hydrophobic ...


    pub_type: 杂志文章


    authors: Kilhoffer MC,Gérard D

    更新日期:1985-09-24 00:00:00

  • Characterization of autocatalytic conversion of precursor BACE1 by heteronuclear NMR spectroscopy.

    abstract::Accumulation of the cytotoxic 40- to 42-residue beta-amyloid peptide represents the primary pathological process in Alzheimer's disease (AD). BACE1 (beta-site APP cleaving enzyme 1) is responsible for the initial required step in the neuronal amyloidogenic processing of beta-amyloid precursor protein and is a major dr...


    pub_type: 杂志文章


    authors: Wang YS,Beyer BM,Senior MM,Wyss DF

    更新日期:2005-12-20 00:00:00

  • A transition state analogue for two pyruvate metabolizing enzymes, lactate dehydrogenase and alanine dehydrogenase.

    abstract::The synthesis of 5-(2-oxalylethyl)-NADH, a reduced nicotinamide adenine dinucleotide (NADH) derivate with pyruvate covalently attached to the 5 position of the dihydronicotinamide ring over an additional methylene group has been described previously (Trommer, W.E., Blume, H., and Kapmeyer, H. (1976) Justus Liebigs Ann...


    pub_type: 杂志文章


    authors: Kapmeyer H,Pfleiderer G,Trommer WE

    更新日期:1976-11-16 00:00:00

  • Three novel oligosaccharides with the sialyl-Lea structure in human milk: isolation by immunoaffinity chromatography.

    abstract::We have determined the structures of three novel oligosaccharides isolated from human milk using the monoclonal antibody MSW 113. These oligosaccharides were purified by affinity chromatography on a column of the immobilized monoclonal antibody and by high-performance liquid chromatography. From the results of 500-MHz...


    pub_type: 杂志文章


    authors: Kitagawa H,Nakada H,Kurosaka A,Hiraiwa N,Numata Y,Fukui S,Funakoshi I,Kawasaki T,Yamashina I,Shimada I

    更新日期:1989-10-31 00:00:00

  • Zn(2+)-dependent misfolding of the p53 DNA binding domain.

    abstract::The DNA binding domain (DBD) of p53 folds by a complex mechanism that involves parallel pathways and multiple intermediates, both on- and off-pathway. This heterogeneity renders DBD particularly susceptible to misfolding and aggregation. The origins of parallel folding mechanisms are not well understood. DBD folding h...


    pub_type: 杂志文章


    authors: Butler JS,Loh SN

    更新日期:2007-03-13 00:00:00

  • Collagen biosynthesis by cultured Chinese hamster lung cells. Cell-free synthesis of procollagen alpha chains.

    abstract::Cell-free extracts from the HTl clone of cultured Chinese hamster lung cells efficiently promote the incorporation of proline into newly synthesized material, 50% of which is digestible to small peptides by highly purified bacterial collagenase. The synthesis of the these products occurs under optimal protein synthesi...


    pub_type: 杂志文章


    authors: Haralson MA,Frey KL,Mitchell WM

    更新日期:1978-03-07 00:00:00