Deletion and purification studies to elucidate the structure of the Actinobacillus actinomycetemcomitans cytolethal distending toxin.

Abstract:

:Cytolethal distending toxin (CDT) is one of the exotoxins produced by Actinobacillus actinomycetemcomitans, an agent of localized aggressive periodontitis. We constructed N-terminal deletion mutants of CdtA using an Escherichia coli expression system and found that ADelta19-47, with a deletion from Asn-19 to Pro-47, showed comparable CDT activity but no apparent heterogeneity of CdtA. The wild-type CDT (wtCDT) and the mutant CDT (ADelta19-47CDT) were purified to homogeneity by introducing a histidine tag into the C-terminal end of CdtB. Both purified wtCDT and purified ADelta19-47CDT showed strong CDT activity and a tripartite structure composed of CdtA (subunit A), 31 kDa CdtB (subunit B), and 18.5 kDa CdtC (subunit C) in nearly a 1:1:1 stoichiometry. Importantly, subunit A was identified as heterogeneous with three CdtA variants in wtCDT, but homogeneous in ADelta19-47CDT. Purified CDTs also showed high stability that was absolutely dependent on the presence of sucrose in the buffer. In conclusion, the region from the Asn-19 to Pro-47 of CdtA contributes to the heterogeneous production of CdtA, but is dispensable for the toxin activity. Furthermore, this study describes an effective protocol for the purification of a native rather than reconstituted CDT, and clarifies the subunit composition of the active CDT holotoxin.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Saiki K,Gomi T,Konishi K

doi

10.1093/jb/mvh121

keywords:

subject

Has Abstract

pub_date

2004-09-01 00:00:00

pages

335-42

issue

3

eissn

0021-924X

issn

1756-2651

pii

136/3/335

journal_volume

136

pub_type

杂志文章
  • Ultrasensitive DNA chip: gene expression profile analysis without RNA amplification.

    abstract::We have developed a new DNA chip whose substrate has a unique minute columnar array structure made of plastic. The DNA chip exhibits ultrahigh sensitivity, up to 100-fold higher than that of reference DNA chips, which makes it possible to monitor gene expression profiles even with very small amounts of RNA (0.1-0.01 m...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvj086

    authors: Nagino K,Nomura O,Takii Y,Myomoto A,Ichikawa M,Nakamura F,Higasa M,Akiyama H,Nobumasa H,Shiojima S,Tsujimoto G

    更新日期:2006-04-01 00:00:00

  • The binding of adenine nucleotides to apo-electron-transferring flavoprotein.

    abstract::Apoprotein of electron-transferring flavoprotein (ETF) reacts with FAD as follows: A*<-->A, A+FAD<-->holoETF. Two different forms of apoETF (A* and A) convert into each other and only one of them, A, can associate with FAD [Sato, K. et al. (1991) J. Biochem. 109, 734-740]. In the present study, the reactions between a...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a123980

    authors: Sato K,Nishina Y,Shiga K

    更新日期:1992-12-01 00:00:00

  • Localization of the phosphatidylserine-binding site of glyceraldehyde-3-phosphate dehydrogenase responsible for membrane fusion.

    abstract::In this study, we demonstrated that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a phosphatidylserine (PS)-binding protein and localized the putative PS-binding site involved in the membrane fusion induced by this enzyme. In an attempt to identify the PS-binding proteins, we raised polyclonal antibodies against...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a021886

    authors: Kaneda M,Takeuchi K,Inoue K,Umeda M

    更新日期:1997-12-01 00:00:00

  • N-terminal amino acid sequence of human urinary prokallikrein.

    abstract::The N-terminal amino acid sequences of human urinary prokallikrein and kallikrein have been determined. Their amino acid sequences are as follows. (Formula; see text) The results showed that prokallikrein comprises an additional seven amino acids at the amino terminus of the kallikrein, of which the sequence is (H2N)A...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a135563

    authors: Takahashi S,Irie A,Katayama Y,Ito K,Miyake Y

    更新日期:1986-03-01 00:00:00

  • Novel oligosaccharides with the sialyl-Le(a) structure in human milk.

    abstract::Two novel oligosaccharides with the sialyl-Le(a) structure were isolated from human milk using a 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, and structurally characterized...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a124207

    authors: Kitagawa H,Nakada H,Fukui S,Funakoshi I,Kawasaki T,Yamashina I,Tate S,Inagaki F

    更新日期:1993-10-01 00:00:00

  • Characterization of the initial steps of precursor import into rat liver mitoplasts.

    abstract::Mitochondria have two independent protein-import machineries, one in the outer membrane (the Tom system) and the other in the inner membrane (the Tim system). Here, we have characterized the initial steps of precursor import into rat liver mitoplasts. The import reaction was separated into two stages, consisting of pr...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a022186

    authors: Ishihara N,Komiya T,Sakaguchi M,Ito A,Mihara K

    更新日期:1998-10-01 00:00:00

  • Mechanism of action of GTP in the induction of Ca2+ release from hepatic microsomes.

    abstract::The mechanism by which GTP induces Ca2+ release from Ca2(+)-preloaded rat hepatic microsomes was studied. In the same concentration range as that for Ca2+ release, GTP inhibited the initial rate of ATP-driven Ca2+ uptake. It also inhibited the formation by ATP of the phosphorylated intermediate of Ca2(+)-ATPase, which...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a123084

    authors: Kimura S,Higham S,Robison BC,Kraus-Friedmann N

    更新日期:1990-04-01 00:00:00

  • Extensible and less-extensible domains of connectin filaments in stretched vertebrate skeletal muscle sarcomeres as detected by immunofluorescence and immunoelectron microscopy using monoclonal antibodies.

    abstract::Two kinds of monoclonal antibodies (3B9 and SM1) against connectin, muscle elastic protein, reacted with both alpha- and beta-connectins. Immunofluorescence studies revealed that 3B9 stained both edges of the A band of chicken breast muscle myofibrils and remained as such upon stretching to a sarcomere length of 3.5 m...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a122499

    authors: Itoh Y,Suzuki T,Kimura S,Ohashi K,Higuchi H,Sawada H,Shimizu T,Shibata M,Maruyama K

    更新日期:1988-10-01 00:00:00

  • Molecular cloning and characterization of two zebrafish alpha(1,3)fucosyltransferase genes developmentally regulated in embryogenesis.

    abstract::Some alpha(1,3)fucosylated oligosaccharides serve as counter receptors to lectin-like adhesion proteins or are expressed with temporal precision during embryogenesis, and alpha(1, 3)fucosyltransferase is a key enzyme in the production of these oligosaccharides. Two alpha(1,3)-fucosyltransferase genes, designated zFT1 ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a022357

    authors: Kageyama N,Natsuka S,Hase S

    更新日期:1999-04-01 00:00:00

  • Structural study of the carbohydrate moiety of bovine pancreatic ribonuclease B.

    abstract::The sugar chain of bovine pancreatic ribonuclease B was released from the polypeptide moiety by hydrazinolysis and by endo-beta-N-acetylglucosaminidase H digestion, and reduced with NaB[3H]4 after N-acetylation. The radioactive oligosaccharide mixtures thus obtained were fractionated by Bio-Gel P-4 column chromatograp...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:

    authors: Liang CJ,Yamashita K,Kobata A

    更新日期:1980-07-01 00:00:00

  • A neutral subtilopeptidase inhibitor from porcine serum some evidence for alpha2-macroglobulin.

    abstract::An inhibitor of neutral subtilopeptidase [EC 3.4.24.4] was purified from porcine serum by salting out with (NH4)2SO4, chromatography on anion exchange sephadex, gel filtration with Sepharose 6B, and isoelectric focusing. The preparation was homogeneous by electrophoretic and ultracentrifugal criteria, and was shown to...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:

    authors: Tsuru D,Tomimatsu M,Fujiwara K,Kawahara K

    更新日期:1975-06-01 00:00:00

  • Metabolism of trichothecene mycotoxins. II. Substrate specificity of microsomal deacetylation of trichothecenes.

    abstract::The substrate specificity of microsomal nonspecific carboxyesterase [EC 3.1.1.1] from rabbit and rat livers was studied in vitro by using seven (A)-type and six (B)-type 12,13-epoxytrichothecene mycotoxins. The C-4 acetyl residues of diacetoxyscirpenol, T-2 toxin, monoacetylnivalenol (fusarenon-X), and diacetylnivalen...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a132175

    authors: Ohta M,Matsumoto H,Ishii K,Ueno Y

    更新日期:1978-09-01 00:00:00

  • Inhibition of rat liver rhodanese by di-, tricarboxylic, and alpha-keto acids.

    abstract::Rat liver rhodanese [EC 2.8.1.1] purified by ammonium sulfate fractionation, CM-cellulose and Sephadex G-200 chromatography yielded two active fractions (I & II). Their molecular weights were estimated to be 1.75 X 10(4) (I) and 1.26 X 10(4) (II) by the gel filtration method. Kinetic studies revealed that Fraction I r...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a130973

    authors: Oi S

    更新日期:1975-10-01 00:00:00

  • The FOXP2 forkhead domain binds to a variety of DNA sequences with different rates and affinities.

    abstract::FOXP2 is a member of the P subfamily of FOX transcription factors, the DNA-binding domain of which is the winged helix forkhead domain (FHD). In this work we show that the FOXP2 FHD is able to bind to various DNA sequences, including a novel sequence identified in this work, with different affinities and rates as dete...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvx003

    authors: Webb H,Steeb O,Blane A,Rotherham L,Aron S,Machanick P,Dirr H,Fanucchi S

    更新日期:2017-07-01 00:00:00

  • Directed evolution of thermostable kanamycin-resistance gene: a convenient selection marker for Thermus thermophilus.

    abstract::The whole-genome sequencing of an extreme thermophile, Thermus thermophilus, is now in progress. Like other genome projects, major concern is shifting from the sequence itself to post-sequencing research such as functional or structural genomics. Under such circumstances, the demand for convenient genetic-engineering ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a022539

    authors: Hoseki J,Yano T,Koyama Y,Kuramitsu S,Kagamiyama H

    更新日期:1999-11-01 00:00:00

  • New gene cluster for lantibiotic streptin possibly involved in streptolysin S formation.

    abstract::Streptolysin S (SLS) is a serum-extractable and oxygen-stable hemolysin produced by Group A Streptococcus. A SLS-deficient mutant in which transposon Tn 916 was inserted in a locus distinct from the sag gene cluster [Nizet et al. (2000) Infect. Immun. 68, 4245-4254] was obtained by filter mating of the transposon-harb...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a002918

    authors: Karaya K,Shimizu T,Taketo A

    更新日期:2001-05-01 00:00:00

  • Orientation of the rhodopsin sugar moiety in bovine disk membrane.

    abstract::Rhodopsin from the bovine rod outer segment contains a covalently linked carbohydrate moiety (Heller, J. & Lawrence, M.A. (1973) Biochemistry 9, 864--868). We studied the location of this carbohydrate moiety on the disk membrane by using ferritin-conjugated concanavalin A and concanavalin A labelled with fluorescein i...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:

    authors: Nakano T,Ikai A,Nishigai M,Noda H

    更新日期:1979-05-01 00:00:00

  • Enhancement of the frequency of initiation by a stimulatory protein of RNA polymerase II.

    abstract::By use of purified RNA polymerase II, it was demonstrated that S-II, a stimulatory protein of RNA polymerase II, enhanced the frequency of initiation of transcription from discrete sites on the promoter region of the silk fibroin gene integrated in a supercoiled plasmid DNA in the presence of manganese. In the absence...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a135020

    authors: Sekimizu K,Horikoshi M,Natori S

    更新日期:1984-12-01 00:00:00

  • Structural and functional characterization of herpes simplex virus 1 immediate-early protein infected-cell protein 22.

    abstract::Of the five HSV1 immediate-early proteins, infected-cell protein 22 (ICP22), the product of the Us1 gene, is a member whose function is less understood. In order to promote better understanding of the role of ICP22 in viral replication, mutation and fluorescence techniques were used to investigate the biochemical rela...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvj135

    authors: Cun W,Hong M,Liu LD,Dong CH,Luo J,Li QH

    更新日期:2006-07-01 00:00:00

  • Isolation of N omega-phosphoarginine hydrolase from rat liver and its physical properties.

    abstract::N omega-Phosphoarginine hydrolase from rat liver cytosol was purified to apparent homogeneity on SDS-PAGE, by employing column chromatographies on Sephadex G-75, DEAE-cellulose, QAE-Toyopearl, and glutathione-2-pyridyl-disulfide-Superose. One milligram protein of the final preparation released 4 mumol/min of inorganic...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a124032

    authors: Yokoyama K,Ohmori H,Kumon A

    更新日期:1993-02-01 00:00:00

  • Presence of 1-O-alk-1'-enyl-2-O-acetyl glycerophosphocholine (vinyl form of PAF) in perfused rat and guinea pig hearts.

    abstract::1-O-Alk-1'-enyl-2-O-acetyl-glycerophosphocholine (vinyl form of PAF) was found with PAF in perfused rat and guinea pig hearts. The main molecular species of the vinyl form of PAF, after separation by reverse phase HPLC, were identified as 1-O-hexadec-, -octadec-, and -octadecen-1'-enyl-2-O-acetyl-GPCs (16:0, 18:0, and...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a122693

    authors: Nakayama R,Saito K

    更新日期:1989-04-01 00:00:00

  • Irradiation with ultraviolet light in the presence of vanadate increases Ca2+ permeability of the sarcoplasmic reticulum membrane via Ca(2+)-ATPase.

    abstract::The sarcoplasmic reticulum (SR) of rabbit skeletal muscle was irradiated with ultraviolet light (UV) in the presence of vanadate plus 2 mM EGTA, 10 mM MgCl2, 20% DMSO, and 50 mM PIPES (pH 6.5) at room temperature. In the presence of 100 microM vanadate, the Ca(2+)-uptake activity of SR rapidly decreased and was almost...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/117.2.324

    authors: Hirose T,Yamasaki K,Yamamoto T

    更新日期:1995-02-01 00:00:00

  • Blocking effect of trypsin associated with ovomucoid on the antibody binding to ovomucoid.

    abstract::Ovomucoid-trypsin association complex was prepared by incubating chicken egg white ovomucoid with bovine trypsin. The reactivity of ovomucoid-trypsin complex was investigated by immunodiffusion, quantitative precipitation and enzyme-linked immunosorbent assay (ELISA). It was demonstrated that the association of trypsi...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a134352

    authors: Matsuda T,Watanabe K,Nakamura R

    更新日期:1983-07-01 00:00:00

  • Characterization of recombinant YakC of Schizosaccharomyces pombe showing YakC defines a new family of aldo-keto reductases.

    abstract::The yakC gene in Schizosaccharomyces pombe, which encodes yakC protein (YakC), a potential member of an aldo-keto reductase (AKR) family, was cloned and expressed in Escherichia coli cells. The recombinant YakC purified to homogeneity catalyzed the reduction of 2-nitrobenzaldehyde (k(cat), 44.1 s(-1), K(m), 0.185 +/- ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a003267

    authors: Morita T,Huruta T,Ashiuchi M,Yagi T

    更新日期:2002-10-01 00:00:00

  • The effects of mutations at position 253 on the thermostability of the Bacillus subtilis 3-isopropylmalate dehydrogenase subunit interface.

    abstract::3-Isopropylmalate dehydrogenase (IPMDH) is a dimeric enzyme with a strongly hydrophobic core that is composed of residues from four alpha-helices. We replaced Glu253, which is found in the hydrophobic core and is part of the subunit interface of the Bacillus subtilis (Bs) IPMDH, with several other amino acids to probe...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvm082

    authors: Ohkuri T,Yamagishi A

    更新日期:2007-06-01 00:00:00

  • Thermotropic transition in the states of proteins in sarcoplasmic reticulum vesicles.

    abstract::Interaction between protein and phospholipid molecules in sarcoplasmic reticulum (SR) was studied by measurement of the conformational fluctuation of the protein using the kinetics of the hydrogen-deuterium exchange reaction. It was revealed that the state of SR ATPase undergoes a phase transition at about 18 degrees ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a131793

    authors: Kirino Y,Anzai K,Shimizu H,Ohta S,Nakanishi M,Tsuboi M

    更新日期:1977-10-01 00:00:00

  • Blood group A activities of glycoprotein and glycolipid from human erythrocyte membranes.

    abstract::It is known that ABO blood group substances in human erythrocyte membranes are sphingoglycolipids, but recently several authors have reported that the glycoproteins of the erythrocyte membranes also have ABO blood group activities in addition to MN blood group activities and virus hemagglutination inhibitor activity. ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a131018

    authors: Yamato K,Handa S,Yamakawa T

    更新日期:1975-12-01 00:00:00

  • Cell surface chondroitin sulphate proteoglycan 4 (CSPG4) binds to the basement membrane heparan sulphate proteoglycan, perlecan, and is involved in cell adhesion.

    abstract::Chondroitin sulphate proteoglycan 4 (CSPG4) is a cell surface proteoglycan highly expressed by tumour, perivascular and oligodendrocyte cells and known to be involved cell adhesion and migration. This study showed that CSPG4 was present as a proteoglycan on the cell surface of two melanoma cell lines, MM200 and Me1007...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvy008

    authors: Tang F,Lord MS,Stallcup WB,Whitelock JM

    更新日期:2018-05-01 00:00:00

  • Increased acid ceramidase expression depends on upregulation of androgen-dependent deubiquitinases, USP2, in a human prostate cancer cell line, LNCaP.

    abstract::Acid ceramidase (ACDase) metabolizes ceramide to sphingosine, leading to sphingosine 1-phosphate production. Reportedly, ACDase has been upregulated in prostate cancer. However, its regulatory mechanism remains unclear. LNCaP (androgen-sensitive prostate cancer cell line) but not PC3 and DU-145, (androgen-unresponsive...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvv039

    authors: Mizutani N,Inoue M,Omori Y,Ito H,Tamiya-Koizumi K,Takagi A,Kojima T,Nakamura M,Iwaki S,Nakatochi M,Suzuki M,Nozawa Y,Murate T

    更新日期:2015-10-01 00:00:00

  • Complexes of myosin subfragment 1 with pyrophosphate and with adenosine diphosphate as studied by phosphorus-31 nuclear magnetic resonance.

    abstract::Myosin subfragment-1.pyrophosphate (S1.PPi) complex and S1.ADP complex were observed with 31P NMR at various temperatures between 0 and 25 degrees C. The signal of S1.PPi complex showed a small temperature dependence, indicating that a single conformation exists for the complex. It also showed that the electron densit...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a123996

    authors: Tanokura M,Ebashi S

    更新日期:1993-01-01 00:00:00