pH dependence studies provide insight into the structure and mechanism of thimet oligopeptidase (EC 3.4.24.15).

Abstract:

:Thimet oligopeptidase (EC 3.4.24.15; TOP) is a Zn(II) endopeptidase implicated in physiological regulation of processes involving neuropeptides. The present study clarifies the active site structure and mechanism of catalysis of TOP. The enzyme exhibited a bell-shaped pH dependence of activity having an acidic limb due to a protonation event with a pK(a) of 5.7 and a basic limb with pK(a) of 8.8. The acidic limb can be attributed to protonation of a residue affecting k(cat) while the alkaline limb may be due to conformational change. Mutation of Tyr612 to Phe resulted in more than 400-fold decrease in activity. This result, supported by modeling studies, implicates Tyr612 in transition state stabilization analogous to the role of His231 of thermolysin.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Sigman JA,Edwards SR,Pabon A,Glucksman MJ,Wolfson AJ

doi

10.1016/s0014-5793(03)00548-9

keywords:

subject

Has Abstract

pub_date

2003-06-19 00:00:00

pages

224-8

issue

2-3

eissn

0014-5793

issn

1873-3468

pii

S0014579303005489

journal_volume

545

pub_type

杂志文章
  • In vitro studies on origin and site of action of enzyme activity responsible for conversion of human proapoprotein A-I into apoprotein A-I.

    abstract::Human hepatocellular carcinoma cells (Hep G2) were shown to secrete apo A-I as a proprotein . No apo A-I synthesis could be detected with endothelial cells from human umbilical cord veins. Conversion of proapo A-I into apo A-I is a slow (of the order of hours) process, mediated by a Ca2+/Mg2+-dependent enzyme which is...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(84)81379-4

    authors: Kooistra T,van Hinsbergh V,Havekes L,Kempen HJ

    更新日期:1984-05-07 00:00:00

  • Human interferon-gamma lacking 23 COOH-terminal amino acids is biologically active.

    abstract::We constructed five mutated cDNAs encoding human interferon-gamma (IFN-gamma) derivatives lacking 19-23 COOH-terminal residues and expressed them in Escherichia coli. All the derivatives were purified to homogeneity. They showed substantially the same order of antiviral activity in vitro as the intact molecule, and be...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(88)80671-9

    authors: Sakaguchi M,Honda S,Ozawa M,Nishimura O

    更新日期:1988-03-28 00:00:00

  • Using an in vivo phagemid system to identify non-compatible loxP sequences.

    abstract::The site-specific recombination system of bacteriophage P1 is composed of the Cre recombinase that recognizes a 34-bp loxP site. The Cre/loxP system has been extensively used to manipulate eukaryotic genomes for functional genomic investigations. The creation of additional heterologous loxP sequences potentially expan...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(01)02541-8

    authors: Siegel RW,Jain R,Bradbury A

    更新日期:2001-06-15 00:00:00

  • Metabolic profiling using combined GC-MS and LC-MS provides a systems understanding of aristolochic acid-induced nephrotoxicity in rat.

    abstract::We present here a combined GC-MS and LC-MS metabolic profiling approach to unraveling the pathological outcomes of aristolochic acid (AA)-induced nephrotoxicity. Urine samples were analyzed by GC-MS and LC-MS in combination with pattern recognition techniques, e.g. principal component analysis (PCA), orthogonal projec...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2007.01.036

    authors: Ni Y,Su M,Qiu Y,Chen M,Liu Y,Zhao A,Jia W

    更新日期:2007-02-20 00:00:00

  • Requirement of tyrosylprotein sulfotransferase-A for proper cuticle formation in the nematode C. elegans.

    abstract::Tyrosine O-sulfation is one of the post-translational modification processes that occur to membrane proteins and secreted proteins in eukaryotes. Tyrosylprotein sulfotransferase (TPST) is responsible for this modification, and in this report, we describe the expression pattern and the biological role of TPST-A in the ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2004.11.044

    authors: Kim TH,Hwang SB,Jeong PY,Lee J,Cho JW

    更新日期:2005-01-03 00:00:00

  • Mode of action of bottromycin A2: effect of bottromycin A2 on polysomes.

    abstract::When bottromycin A2 was added to an in vitro protein synthesis system carried out by naturally occurring polysomes, it inhibited protein synthesis effectively. Examination of the 3 steps of peptide chain elongation revealed that the binding of aminoacyl-tRNA to the polyribosomes was inhibited by bottromycin A2. In con...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(83)80118-5

    authors: Otaka T,Kaji A

    更新日期:1983-03-07 00:00:00

  • Coupling of D2 and D3 dopamine receptors to G-proteins.

    abstract::Recombinant cell lines expressing D2 and D3 dopamine receptor isoforms have been used to study coupling of these receptors to G-proteins from the effects of GTP to reduce agonist binding affinities in agonist/[3H]spiperone competition experiments. D2(short) expressed in Ltk- cells, D2(long) expressed in Ltk- or CHO ce...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(93)81168-y

    authors: Castro SW,Strange PG

    更新日期:1993-01-11 00:00:00

  • Biosynthetic lipid-tagging of antibodies.

    abstract::Chemical conjugation of fatty acids to antibodies generates lipid-modified molecules which have found use in the targeting of liposome-mediated drug delivery and in liposome-based immunoassays. Alternatively, bacterial expression of antibodies as single-chain FV fragments fused to lipoprotein signal peptide and N-term...

    journal_title:FEBS letters

    pub_type: 杂志文章,评审

    doi:10.1016/0014-5793(94)00313-0

    authors: Keinänen K,Laukkanen ML

    更新日期:1994-06-06 00:00:00

  • An in vivo assay for the identification of target proteases which cleave membrane-associated substrates.

    abstract::Proteases not only play a fundamental role in numerous physiological processes, but are also involved in several human diseases including Alzheimer's disease (AD). A key protease implicated in AD is the so far unidentified gamma-secretase, which cleaves the membrane-bound beta-amyloid precursor protein (betaAPP) at th...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(99)01627-0

    authors: Steiner H,Pesold B,Haass C

    更新日期:1999-12-17 00:00:00

  • Regulation of the MAP kinase cascade in PC12 cells: B-Raf activates MEK-1 (MAP kinase or ERK kinase) and is inhibited by cAMP.

    abstract::In PC12 cells, cAMP stimulates the MAP kinase pathway by an unknown mechanism. Firstly, we examined the role of calcium ion mobilization and of protein kinase C in cAMP-stimulated MAP kinase activation. We show that cAMP stimulates p44mapk independently of these events. Secondly, we studied the role of B-Raf in this p...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(94)01376-c

    authors: Peraldi P,Frödin M,Barnier JV,Calleja V,Scimeca JC,Filloux C,Calothy G,Van Obberghen E

    更新日期:1995-01-09 00:00:00

  • Rat glutathione transferase 8-8, an enzyme efficiently detoxifying 4-hydroxyalk-2-enals.

    abstract::Rat glutathione transferase 8-8 is one of the less abundant cytosolic glutathione transferases, accounting for approx. 1% of the total activity with 1-chloro-2,4-dinitrobenzene in liver. The enzyme is eluted at pH 6.3 upon chromatofocusing and has so far been identified in liver, kidney, lung and testis. Characteristi...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(86)80743-8

    authors: Jensson H,Guthenberg C,Alin P,Mannervik B

    更新日期:1986-07-28 00:00:00

  • Functional analysis of an indole-diterpene gene cluster for lolitrem B biosynthesis in the grass endosymbiont Epichloë festucae.

    abstract::Epichloë festucae Fl1 in association with Lolium perenne synthesizes a diverse range of indole-diterpene bioprotective metabolites, including lolitrem B, a potent tremorgen. The ltm genes responsible for the synthesis of these metabolites are organized in three clusters at a single sub-telomeric locus in the genome of...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2012.06.035

    authors: Saikia S,Takemoto D,Tapper BA,Lane GA,Fraser K,Scott B

    更新日期:2012-07-30 00:00:00

  • The role of mild uncoupling and non-coupled respiration in the regulation of hydrogen peroxide generation by plant mitochondria.

    abstract::The roles of mild uncoupling caused by free fatty acids (mediated by plant uncoupling mitochondrial protein (PUMP) and ATP/ADP carrier (AAC)) and non-coupled respiration (alternative oxidase (AO)) on H(2)O(2) formation by plant mitochondria were examined. Both laurate and oleate prevent H(2)O(2) formation dependent on...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(00)01576-3

    authors: Casolo V,Braidot E,Chiandussi E,Macrì F,Vianello A

    更新日期:2000-05-26 00:00:00

  • Hydrolyse des aminoacyl-tARN par la phosphodiesterase du venin.

    abstract::The influence of the esterification of the 3'(2') hydroxyl group of tRNA on the hydrolysis by venom phosphodiesterase (Crotalus adamanteus) was studied. It was shown that this esterification does not change the rate of hydrolysis. At pH 7-8, no significant differences were observed either when the alpha amino group of...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(68)80046-8

    authors: Yot P,Gueguen P,Chapeville F

    更新日期:1968-08-01 00:00:00

  • Interaction of nitrogen dioxide with human plasma. Antioxidant depletion and oxidative damage.

    abstract::Nitrogen dioxide (NO2.) is often present in inhaled air and may be generated in vivo from nitric oxide. Exposure of human blood plasma to NO2. caused rapid losses of ascorbic acid, uric acid and protein thiol groups, as well as lipid peroxidation and depletions of alpha-tocopherol, bilirubin and ubiquinol-10. No incre...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(92)81185-o

    authors: Halliwell B,Hu ML,Louie S,Duvall TR,Tarkington BK,Motchnik P,Cross CE

    更新日期:1992-11-16 00:00:00

  • A possible role of exon-shuffling in the evolution of signal peptides of human proteins.

    abstract::It was recently shown that there is a predominance of phase 1 introns near the cleavage site of signal peptides encoded by human genes. It was suggested that this biased distribution was due to intron insertion at AGmid R:G proto-splice sites. However, we found that there is no disproportional excess of AGmid R:G that...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2006.01.094

    authors: Vibranovski MD,Sakabe NJ,de Souza SJ

    更新日期:2006-03-06 00:00:00

  • The fMet-tRNA binding domain of translational initiation factor IF2: role and environment of its two Cys residues.

    abstract::Mutations of the cysteines (positions 668 and 714) were generated in the IF2 C domain of Bacillus stearothermophilus translation initiation factor IF2. The corresponding proteins were characterized functionally and structurally. Most (yet not all) amino acid replacements at both positions resulted in severe reduction ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(99)01280-6

    authors: Misselwitz R,Welfle K,Krafft C,Welfle H,Brandi L,Caserta E,Gualerzi CO

    更新日期:1999-10-15 00:00:00

  • Synthesis and absolute configuration of optically pure enantiomers of a kappa-opioid receptor selective agonist.

    abstract::The enantiomers of U50,488, ligands highly selective for kappa-opioid receptors, have been prepared by a refined procedure and their optical purity demonstrated. The absolute configuration of (+)-trans-2-pyrrolidinyl-N-methylcyclohexylamine, a chemically versatile intermediate for synthesis of analogs of kappa-opioid ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(87)80315-0

    authors: DeCosta B,George C,Rothman RB,Jacobson AE,Rice KC

    更新日期:1987-11-02 00:00:00

  • The structure of procalcitonin of the salmon as deduced from its cDNA sequence.

    abstract::Oligonucleotide probes based on the known amino acid sequence of salmon calcitonin were used to screen a cDNA library obtained from ultimobranchial glands of salmon for clones encoding salmon calcitonin. From the cDNA sequence of strongly hybridizing clones the complete primary structure of the calcitonin precursor co...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(87)80558-6

    authors: Pöschl E,Lindley I,Hofer E,Seifert JM,Brunowsky W,Besemer J

    更新日期:1987-12-21 00:00:00

  • A kinome of 2600 in the ciliate Paramecium tetraurelia.

    abstract::Protein kinases play a crucial role in the regulation of cellular processes. Most eukaryotes reserve about 2.5% of their genes for protein kinases. We analysed the genome of the single-celled ciliate Paramecium tetraurelia and identified 2606 kinases, about 6.6% of its genes, representing the largest kinome to date. A...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2009.10.029

    authors: Bemm F,Schwarz R,Förster F,Schultz J

    更新日期:2009-11-19 00:00:00

  • Identification of a novel protein complex containing annexin A4, rabphilin and synaptotagmin.

    abstract::Rabphilin is a synaptic vesicle-associated protein proposed to play a role in regulating neurotransmitter release. Here we report the isolation and identification of a novel protein complex containing rabphilin, annexin A4 and synaptotagmin 1. We show that the rabphilin C2B domain interacts directly with the N-terminu...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/S0014-5793(03)01513-8

    authors: Willshaw A,Grant K,Yan J,Rockliffe N,Ambavarapu S,Burdyga G,Varro A,Fukuoka S,Gawler D

    更新日期:2004-02-13 00:00:00

  • Accumulation of protoporphyrin IX in light-sensitive mutants of Escherichia coli.

    abstract::The accumulation of protoporphyrin IX (Proto IX) in light-sensitive mutants of Escherichia coli was detected by spectrofluorimetry. Fluorescence emission and excitation spectra were recorded from extracts of bacterial cells. Proto IX clearly accumulated in cells with mutations in the visA (hemH) gene but not in the wi...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(92)81341-i

    authors: Miyamoto K,Nishimura K,Masuda T,Tsuji H,Inokuchi H

    更新日期:1992-10-05 00:00:00

  • Purification of a heat-stable activator protein for ADP-ribosylation factor-dependent phospholipase D.

    abstract::A heat-stable activator for ADP-ribosylation factor (ARF)-dependent phospholipase D (PLD) was purified to near homogeneity from rat kidney cytosol by a sequential column chromatography. The purified activator has a molecular mass of 23 kDa on SDS-PAGE. Using a partially purified ARF-dependent PLD from rat kidney, the ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(97)01611-6

    authors: Akisue T,Jinnai H,Hitomi T,Miwa N,Yoshida K,Nakamura S

    更新日期:1998-01-23 00:00:00

  • Suppression of a phosphatidylinositol 3-kinase signal by a specific spliced variant of Drosophila PTEN.

    abstract::Drosophila PTEN (dPTEN) plays indispensable roles in the development of Drosophila melanogaster by controlling cell size and number. Although three potential spliced forms of dPTEN have been isolated, functional distinction among these forms remains elusive. In this study, we demonstrate that all spliced forms of dPTE...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2004.03.074

    authors: Maehama T,Kosaka N,Okahara F,Takeuchi K,Umeda M,Dixon JE,Kanaho Y

    更新日期:2004-05-07 00:00:00

  • OSBPa, a predicted oxysterol binding protein of Dictyostelium, is required for regulated entry into culmination.

    abstract::The oxysterol binding proteins (OSBPs) are believed to control cholesterol homeostasis but their precise mechanism of action is not well understood. The Dictyostelium osbA gene encodes a predicted OSBP, OSBPa, which lacks the PH domain that in most other OSBPs directs targetting to the Golgi. OSBPa instead localises s...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(02)03150-2

    authors: Fukuzawa M,Williams JG

    更新日期:2002-09-11 00:00:00

  • Studies on the interaction of the dye, stains-all, with individual calcium-binding domains of calmodulin.

    abstract::We show that the calcium-mimic dye, Stains-all, is a convenient probe to study the structural features of the individual calcium-binding sites of calmodulin (CaM) and related calcium-binding proteins (CaBP). These peptides bind the dye in their calcium-binding sites, and induce a circular dichroism (CD) band in the bo...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(93)81761-n

    authors: Sharma Y,Gopalakrishna A,Balasubramanian D,Fairwell T,Krishna G

    更新日期:1993-07-12 00:00:00

  • Light-induced stimulation of carbonic anhydrase activity in pea thylakoids.

    abstract::Stimulation of the bicarbonate dehydration reaction in thylakoid suspension under conditions of saturating light at pH 7.6-8.0 was discovered. This effect was inhibited by nigericin or the lipophilic carbonic anhydrase (CA) inhibitor ethoxyzolamide (EZ), but not by the hydrophilic CA inhibitor, acetazolamide. It was s...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(00)01328-4

    authors: Moskvin OV,Ivanov BN,Ignatova LK,Kollmeier MA

    更新日期:2000-03-31 00:00:00

  • A human B-box-binding protein downregulated in adenovirus 5-transformed human cells.

    abstract::Internal promoters of some genes transcribed by RNA polymerase III (e.g. tRNA genes, adenovirus VA1 RNA gene, human retroposons of the Alu family) contain a conserved sequence element, B-box, interacting with basal transcription factor TFIIIC2 which initiates assembly of the full transcription complex on the genes, an...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(96)00353-5

    authors: Kropotov AV,Tomilin NV

    更新日期:1996-05-13 00:00:00

  • Structural and functional characterization of the bacterial translocation inhibitor GE82832.

    abstract::The structure of GE82832, a translocation inhibitor produced by a soil microorganism, is shown to be highly related to that of dityromycin, a bicyclodecadepsipeptide antibiotic discovered long ago whose characterization had never been pursued beyond its structural elucidation. GE82832 and dityromycin were shown to int...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2012.07.040

    authors: Brandi L,Maffioli S,Donadio S,Quaglia F,Sette M,Milón P,Gualerzi CO,Fabbretti A

    更新日期:2012-09-21 00:00:00

  • Long-term regulation of voltage-gated Ca(2+) channels by gabapentin.

    abstract::Gabapentin (GBP) is a gamma-aminobutyric acid analog effective in the treatment of seizures. A high-affinity interaction between GBP and the alpha(2)delta subunit of the voltage-gated Ca(2+) channels has been documented. In this report, we examined the effects of the chronic treatment with GBP on neuronal recombinant ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(02)03295-7

    authors: Kang MG,Felix R,Campbell KP

    更新日期:2002-09-25 00:00:00