Effects of salts on thermolysin: activation of hydrolysis and synthesis of N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester, and a unique change in the absorption spectrum of thermolysin.

Abstract:

:It has been reported that neutral salts such as NaCl activate the thermolysin-catalyzed hydrolysis of substrates containing glycine at the P1 position (carboxylic side of the cleavage bond) [Holmquist, B. & Vallee, B.L. (1976) Biochemistry 15, 101-107]. In this paper, we demonstrate that high concentrations (1-4 M) of neutral salts greatly enhance the thermolysin activity in both hydrolysis and synthesis of N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester (ZAPM), a precursor of a peptide sweetener, aspartame, in which the L-aspartyl residue is the P1 residue. The enzyme activity is enhanced with an increase in salt concentration in a pseudo-exponential fashion. The degree of activation by salts was in the order LiCl > NaCl > KCl. The rate of ZAPM hydrolysis in the presence of 3.8 M NaCl was 6-7 times higher than that in its absence, and 50 times or more activation is expected in saturated NaCl solution. The activation is brought about solely through an increase in the catalytic constant (kcat), and the Michaelis constant (Km) is not affected at all by the presence of NaCl. On mixing thermolysin with NaCl, a unique absorption difference spectrum suggesting a conformational change of the enzyme was observed. The intensity increased in a pseudo-exponential fashion with increase of NaCl concentration up to 3 M, and this dependence is similar to that of the enzyme activity.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Inouye K

doi

10.1093/oxfordjournals.jbchem.a123901

keywords:

subject

Has Abstract

pub_date

1992-09-01 00:00:00

pages

335-40

issue

3

eissn

0021-924X

issn

1756-2651

journal_volume

112

pub_type

杂志文章
  • Proton-relay system of carboxypeptidase Y as a sole catalytic site: studies on mutagenic replacement of his 397.

    abstract::His397 was replaced with alanine by site-directed mutagenesis of the cloned PRC1 gene in order to confirm the role of this residue in the proton-relay system of carboxypeptidase Y (CPY). The expressed and purified H397A showed a CD spectrum almost identical to that of the wild type enzyme, but its heat stability and c...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a022133

    authors: Jung G,Ueno H,Hayashi R

    更新日期:1998-08-01 00:00:00

  • Cystatin S: a cysteine proteinase inhibitor of human saliva.

    abstract::An acidic protein of human saliva, which we named SAP-1 previously, is now shown to be an inhibitor of several cysteine proteinases. The protein inhibited papain and ficin strongly, and stem bromelain and bovine cathepsin C partially. However, it did not inhibit either porcine cathepsin B or clostripain. The mode of t...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a134952

    authors: Isemura S,Saitoh E,Ito S,Isemura M,Sanada K

    更新日期:1984-10-01 00:00:00

  • Ribonuclease H from rat liver. II. Partial purification and characterization of cytosol ribonuclease H1.

    abstract::We have detected in rat liver cytosol three enzymes (termed C-1, C-2, and C-3) which cleaved the RNA moiety of RNA-DNA hybrid. These enzymes were separated from each other by DEAE-Sephadex and Sephadex G-200 chromatography. C-1 and C-2 specifically act on the RNA moiety of RNA-DNA hybrid, while C-3 degrades single-str...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a132140

    authors: Tashiro F,Ueno Y

    更新日期:1978-08-01 00:00:00

  • Subcellular distribution of GTP-binding proteins, Go and Gi2, in rat cerebral cortex.

    abstract::The localization of GTP-binding protein (G-protein) subunits, Go alpha, Gi2 alpha and beta, in subcellular fractions of rat cerebral cortex was determined by means of immunoassays specific for the respective subunits. High concentrations of all three subunits were observed in both crude mitochondrial and microsomal fr...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a123110

    authors: Asano T,Nagahama M,Kato K

    更新日期:1990-05-01 00:00:00

  • Lysophospholipase L1 from Escherichia coli K-12 overproducer.

    abstract::After screening 900 E. coli strains of the Clarke and Carbon collection for by lysophospholipase L1 activities, we isolated a clone bearing the plasmid pLC6-34, which showed an increased level of lysophospholipase L1 activity. Strains bearing the plasmid pC124, a subclone of pLC6-34 in plasmid vector pUC8, showed appr...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:

    authors: Karasawa K,Kudo I,Kobayashi T,Homma H,Chiba N,Mizushima H,Inoue K,Nojima S

    更新日期:1991-02-01 00:00:00

  • Fractionation, isolation and characterization of nonhistone chromosomal protein from calf thymus.

    abstract::1) A method is described for the separation and fractionation of nonhistone chromosomal proteins from salt-urea dissociated calf thymus chromatin. After precipitating DNA in the dissociated chromatin solution with LaCl3, the chromosomal proteins in the supernatant were fractionated by SP-Sephadex C-25 column chromatog...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a130800

    authors: Yoshida M,Kikuchi A,Shimura K

    更新日期:1975-05-01 00:00:00

  • Purification and enzymatic properties of rat serum carboxylesterase.

    abstract::Rat serum carboxylesterase [carboxylic ester hydrolase, EC 3.1.1.1] was purified by ammonium sulfate precipitation, chromatography on DEAE-cellulose, QAE Sephadex A-50 and brushite, and gel filtration on Sephadex G-200. This purified enzyme was shown to be a single protein band on slab gel electrophoresis and its fina...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a132254

    authors: Hashinotsume M,Higashino K,Hada T,Yamamura Y

    更新日期:1978-12-01 00:00:00

  • Analysis of recognition elements for mitochondrial processing peptidase using artificial amino acids: roles of the intervening portion and proximal arginine.

    abstract::We recently demonstrated, using synthetic peptides modeled on the extension peptide of malate dehydrogenase, that amino acid residues present at the proximal and distal positions relative to the cleavage site are critical determinants for the recognition of substrates by mitochondrial processing peptidase [Niidome et ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a022529

    authors: Moriwaki K,Ogishima T,Ito A

    更新日期:1999-11-01 00:00:00

  • Octamer binding protein-1 is involved in inhibition of inducible nitric oxide synthase expression by exogenous nitric oxide in murine liver cells.

    abstract::Nitric oxide (NO) has diverse effects on immune responses and hepatic functions. In BNL CL.2 cells, the murine embryonic liver cells, inducible nitric oxide synthase (iNOS) mRNA expression appeared after 3 h of treatment with IFN-gamma and LPS. Interestingly, mRNA and protein expression of iNOS was down-regulated by s...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a002839

    authors: Lee BS,Kim YM,Kang HS,Kim HM,Pyun KH,Choi I

    更新日期:2001-01-01 00:00:00

  • Transmembrane signal transduction and osmoregulation in Escherichia coli: functional importance of the transmembrane regions of membrane-located protein kinase, EnvZ.

    abstract::EnvZ is a membrane-located protein kinase which modulates expression of the ompF and ompC genes through phosphotransfer signal transduction in Escherichia coli. Previously, we developed an in vitro method for analyzing the intact form of EnvZ in isolated cytoplasmic membranes, and demonstrated that this particular for...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a123823

    authors: Tokishita S,Kojima A,Mizuno T

    更新日期:1992-06-01 00:00:00

  • Rescuing activity of galactoglycerolipids from cellular lesions induced by 5-aminolevulinic acid.

    abstract::An anti-oxygen radical reagent of a bacterial metabolite, M874 monogalactoglycerolipid (di-O-12-methyl-tetradecanoyl-3-O-beta-D-galactopyranosyl-sn-glycerol ), was tested for its ability to protect two organisms against cellular lesions induced by 5-aminolevulinic acid (ALA) and light. In Corynebacterium flavescens AT...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a022674

    authors: Nakata K

    更新日期:2000-05-01 00:00:00

  • Human erythrocyte bisphosphoglycerate mutase: inactivation by glycation in vivo and in vitro.

    abstract::2,3-Bisphosphoglycerate mutase (BPGM) [EC 5.4.2.4] is a multifunctional enzyme that catalyzes both the synthesis and the degradation of 2,3-diphosphoglycerate (2,3-DPG) and contains three types of activities in that it functions as a 2,3-DPG synthetase, a phosphoglycerate mutase and a 2,3-DPG phosphatase. In humans, B...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a022243

    authors: Fujita T,Suzuki K,Tada T,Yoshihara Y,Hamaoka R,Uchida K,Matuo Y,Sasaki T,Hanafusa T,Taniguchi N

    更新日期:1998-12-01 00:00:00

  • Effect of urea on protein separation by ion-exchange chromatography.

    abstract::Ion-exchange chromatography (IEC) is the most frequently used chromatographic technique for the separation of proteins and peptides. In this article, the effects of urea on IEC separation of kiwifruit actinidin, egg white and urinary proteins were examined. The purity and relative amount of each protein in different c...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvq008

    authors: Khademi F,Mostafaie A

    更新日期:2010-05-01 00:00:00

  • Studies on the ferricytochrome a-ferrocytochrome a3-carbon monoxide complex of mammalian cytochrome oxidase. Conditions for preparation and some properties.

    abstract::The conditions for the preparation of the ferricytochrome a-ferrocytochrome a3-carbon monoxide complex (a3+, a3(2)+CO) of cytochrome oxidase [EC 1.9.3.1] by the ferricyanide-reoxidation method and some properties of the prepared complex were studied. The addition of a small volume of concentrated ferricyanide solution...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a134255

    authors: Horie S,Watanabe T,Ave K

    更新日期:1983-04-01 00:00:00

  • Identification and cloning of the gene coding for lysophospholipase L2 of E. coli K-12.

    abstract::E. coli bearing hybrid plasmid pKOl (Oeda et al. (1981) Mol. Gen. Genet. 184, 191-199) expressed a large amount of lysophospholipase L2 activity. When a mutant which was defective in lysophospholipase L2 activity was transformed with plasmid pKOl, it overproduced lysophospholipase L2 activity. The gene responsible for...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a134566

    authors: Homma H,Kobayashi T,Ito Y,Kudo I,Inoue K,Ikeda H,Sekiguchi M,Nojima S

    更新日期:1983-12-01 00:00:00

  • Methionine aminopeptidase from the hyperthermophilic Archaeon Pyrococcus furiosus: molecular cloning and overexpression in Escherichia coli of the gene, and characteristics of the enzyme.

    abstract::A gene for a methionine aminopeptidase (MAP; EC 3.4.11.18), which catalyzes the removal of amino-terminal methionine from the growing peptide chain on the ribosome, has been cloned from the hyperthermophilic Archaeon, Pyrococcus furiosus, by a novel method effectively using its cosmid protein library, sequenced and ex...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a021831

    authors: Tsunasawa S,Izu Y,Miyagi M,Kato I

    更新日期:1997-10-01 00:00:00

  • Kinetic studies on the hydrolysis of N-acetylated and N-deacetylated derivatives of 4-methylumbelliferyl chitobioside by the family 18 chitinases ChiA and ChiB from Serratia marcescens.

    abstract::Kinetic analyses of the hydrolysis reactions of N-acetylated and N-deacetylated derivatives of 4-methylumbelliferyl chitobioside [(GlcNAc)(2)-UMB (1), GlcN-GlcNAc-UMB (2), GlcNAc-GlcN-UMB (3), and (GlcN)(2)-UMB (4)] by ChiA and ChiB from Serratia marcescens were performed. Both enzymes released UMB from all compounds ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvg031

    authors: Honda Y,Kitaoka M,Tokuyasu K,Sasaki C,Fukamizo T,Hayashi K

    更新日期:2003-02-01 00:00:00

  • Hepatic uptake of bilirubin diglucuronide: analysis by using sinusoidal plasma membrane vesicles.

    abstract::In order to characterize the mechanism for bilirubin transport in the liver, the uptake of bilirubin diglucuronide (BDG) into purified sinusoidal plasma membrane vesicles was investigated. BDG uptake was saturable, and was inhibited by sulfobromophthalein and unconjugated bilirubin, but was not affected by sodium taur...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a123120

    authors: Adachi Y,Roy-Chowdhury J,Roy-Chowdhury N,Kinne R,Tran T,Kobayashi H,Arias IM

    更新日期:1990-05-01 00:00:00

  • Reconstitution of urea-dissociated subunits of a pig heart phosphoprotein phosphatase.

    abstract::Pig heart phosphoprotein phosphatase [phosphoprotein phosphophydrolase, EC 3.1.3.16] of Mr 224,000 was dissociated by gel-filtration on Sephacryl S-300, into an active subunit (alpha subunit) of Mr 31,000 and inactive subunits of higher molecular weight in the presence of 6 M urea. After the removal of urea, these sub...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a133542

    authors: Imazu M,Imaoka T,Usui H,Kinohara N,Takeda M

    更新日期:1981-09-01 00:00:00

  • Signal transduction and sporulation in Bacillus subtilis: heterologous phosphorylation of Spo0A, a sporulation initiation gene product.

    abstract::Spo0A is both a positive and a negative transcriptional regulator which plays a very important role in sporulation initiation in Bacillus subtilis. Its N-terminal amino acid sequence is homologous to that of regulator proteins of the two-component regulatory systems involved in signal transduction in bacteria. Phospho...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a124185

    authors: Asayama M,Kobayashi Y

    更新日期:1993-09-01 00:00:00

  • Crystal structure of adenylate kinase from an extremophilic archaeon Aeropyrum pernix with ATP and AMP.

    abstract::The crystal structure of an adenylate kinase from an extremophilic archaeon Aeropyrum pernix was determined in complex with full ligands, ATP-Mg2+ and AMP, at a resolution of 2.0 Å. The protein forms a trimer as found for other adenylate kinases from archaea. Interestingly, the reacting three atoms, two phosphorus and...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvaa043

    authors: Shibanuma Y,Nemoto N,Yamamoto N,Sampei GI,Kawai G

    更新日期:2020-09-01 00:00:00

  • Two J domains ensure high cochaperone activity of DnaJ, Escherichia coli heat shock protein 40.

    abstract::Heat shock protein 70 (Hsp70) chaperone systems consist of Hsp70, Hsp40 and a nucleotide-exchange factor and function to help unfolded proteins achieve their native conformations. Typical Hsp40s assume a homodimeric structure and have both chaperone and cochaperone activity. The dimeric structure is critical for chape...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvy038

    authors: Uchida T,Kanemori M

    更新日期:2018-08-01 00:00:00

  • A novel fucosylated glycosphingolipid with a Gal beta 1-4Glc beta 1-3Gal sequence in plerocercoids of the parasite, Spirometra erinacei.

    abstract::A novel glycosphingolipid (SEGLx) has been isolated from the plerocercoids of a tapeworm, Spirometra erinacei. From the results of compositional analysis, methylation analysis, exoglycosidase hydrolysis, acid hydrolysis, fast atom bombardment mass spectrometry, and proton nuclear magnetic resonance (NMR) analysis, its...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a124236

    authors: Kawakami Y,Nakamura K,Kojima H,Suzuki M,Inagaki F,Suzuki A,Sonoki S,Uchida A,Murata Y,Tamai Y

    更新日期:1993-11-01 00:00:00

  • Incorporation of serine and ethanolamine into the phospholipids in rabbit retina.

    abstract::The incorporation of serine and ethanolamine into phospholipids in rabbit retinal subcellular fractions and in excised retinas was studied in vitro, and some enzymic properties of the incorporation of phospholipid bases by base exchange were examined in the microsomal fraction. The retina was found to have a higher ra...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a131256

    authors: Mizuno A

    更新日期:1976-07-01 00:00:00

  • Ca2+-binding of S-100 protein in the presence of K+ and Mg2+.

    abstract::Ca2+-binding of S-100 protein was studied using a Ca2+ electrode at pH 6.80. In the presence of 0.1 M KCl and 10 mM MgCl2 (ionic strength 0.13), Ca2+-binding to S-100 protein occurred in three steps with positive cooperativity. The numbers of bound Ca2+ ions in the three steps were 2, 2, and 4. The Ca2+-binding consta...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a122596

    authors: Matsuda S

    更新日期:1988-12-01 00:00:00

  • Separation and properties of three forms of cathepsin H-like cysteine proteinase from rat spleen.

    abstract::Three forms of cathepsin H-like cysteine proteinase were purified from rat spleen by a method involving acid treatment and chromatography on pepstatin-Sepharose, Sephadex G-75, DEAE-Sephacel, CM-Toyopearl, and concanavalin A-Sepharose. The final preparations of these forms all migrated as single protein bands on polya...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a134629

    authors: Yamamoto K,Kamata O,Kato Y

    更新日期:1984-02-01 00:00:00

  • Urinary excretion of retinol-binding protein (RBP) in rabbit chronically poisoned with cadmium. Isolation and partial characterization of rabbit RBP.

    abstract::Studies were conducted to induce urinary excretion of retinol-binding protein (RBP) in the rabbit by long-term administration of cadmium, and to isolate and characterize rabbit RBP. Two rabbits were exposed to Cd at a dose of 0.8 and/or 1.5mg/kg of body weight by means of subcutaneous injections five times per week. O...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a131130

    authors: Muto Y,Nakanishi M,Shidoji Y

    更新日期:1976-04-01 00:00:00

  • Biosynthesis of enzyme-bound formylvaline and formylvalylglycine. A possible initiation complex for gramicidin A biosynthesis.

    abstract::A partially purified fraction (Component I) which catalyzed the ATP-PPi exchange reaction dependent only on valine and glycine among the five constituent amino acids of gramicidin A was obtained from crude extracts of Bacillus brevis ATCC 8185, which produces both gramicidin A and tyrocidines. This fraction was found ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a131446

    authors: Akashi K,Kubota K,Kurahashi K

    更新日期:1977-01-01 00:00:00

  • Further study on association of 5SrRNA-L5 protein complex and methionyl-tRNA to methionyl-tRNA synthetase in the macromolecular aminoacyl-tRNA synthetase complex.

    abstract::To obtain direct evidence for the attachment of 5SrRNA-ribosomal L5 protein particles (5SRNP) and methionine-tRNA (tRNA(met)) to methionyl-tRNA synthetase (MetRS) in the macromolecular aminoacyl-tRNA synthetase (ARS) complex of rat liver, a MetRS-5SRNP-tRNA(met) complex was dissociated from the macromolecular ARS comp...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a021500

    authors: Ogata K,Ohno R,Morioka S,Terao K

    更新日期:1996-11-01 00:00:00

  • Antibacterial activity of peptides derived from the C-terminal region of a hemolytic lectin, CEL-III, from the marine invertebrate Cucumaria echinata.

    abstract::Several synthetic peptides derived from the C-terminal domain sequence of a hemolytic lectin, CEL-III, were examined as to their action on bacteria and artificial lipid membranes. Peptide P332 (KGVIFAKASVSVKVTASLSK-NH(2)), corresponding to the sequence from residue 332, exhibited strong antibacterial activity toward G...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvh007

    authors: Hatakeyama T,Suenaga T,Eto S,Niidome T,Aoyagi H

    更新日期:2004-01-01 00:00:00