Effect of polyethylene glycols on the function and structure of thiol proteases.

Abstract:

:Thiol proteases are industrially significant proteins with catalytic efficiency. The effect of low, medium and high molecular-weight poly (ethylene glycol) (PEG- 400, 6000 and 20000) on the stability of thiol proteases (papain, bromelain and chymopapain) has been studied by activity measurements using synthetic substrate. Structural studies performed on papain by far UV circular dichroism spectroscopic measurements indicate that there is loss in secondary structure of the protein in presence of increasing concentration of PEGs. Intrinsic fluorescence measurements lead us to conclude that tryptophan residues of protein encounter non-polar microenvironment in presence of PEG solvent while acrylamide quenching shows greater accessibility of tryptophan residues of papain in presence of PEGs. Extrinsic fluorescence measurements lead us to conclude that PEGs bind to the hydrophobic sites on the protein and thus destabilize it. Thermal denaturation studies show that melting temperature of papain is decreased in presence of PEGs. Possible mechanism of destabilization is discussed next. The results imply that caution must be exercised in the use of PEGs with thiol proteases or hydrophobic proteins in general, for different industrial applications, even at room temperature.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Fatima S,Khan RH

doi

10.1093/jb/mvm108

subject

Has Abstract

pub_date

2007-07-01 00:00:00

pages

65-72

issue

1

eissn

0021-924X

issn

1756-2651

pii

mvm108

journal_volume

142

pub_type

杂志文章
  • Marked difference between self-aggregations of first and fourth repeat peptides on tau microtubule-binding domain in acidic solution.

    abstract::The heparin-induced self-aggregation behaviours of four repeat peptides (R1-R4) in an acidic solution (pH = 4.5) were investigated by fluorescence and circular dichroism (CD) measurements and compared with those in a neutral solution (pH = 7.5). In contrast with the self-aggregation-resistive behaviours of the R1 and ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvm099

    authors: Mizushima F,Minoura K,Tomoo K,Sumida M,Taniguchi T,Ishida T

    更新日期:2007-07-01 00:00:00

  • Differential rate assay of human pancreatic and salivary alpha-amylases in serum using two coupled enzymes.

    abstract::p-Nitrophenyl O-6-deoxy-6-[(2-pyridyl)amino]-alpha-D-glucopyranosyl-(1----4)-O-alpha- D-glucopyranosyl-(1----4)-O-alpha-D-glucopyranosyl-(1----4)-O-alpha-D- glucopyranosyl-(1----4)-alpha-D-glucopyranoside (FG5P) is hydrolyzed by human pancreatic a-amylase (HPA) or salivary alpha-amylase (HSA) to O-6-deoxy-6-[(2-pyridy...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a121841

    authors: Omichi K,Ikenaka T

    更新日期:1986-11-01 00:00:00

  • Mechanism of inhibition of hepatic triglyceride lipase from human postheparin plasma by apolipoproteins A-I and A-II.

    abstract::The present data describe the mechanism of the inhibitory effects of human plasma apolipoproteins A-I and A-II on hydrolysis of triglyceride catalyzed by hepatic triglyceride lipase using a substrate of triolein particles stabilized with gum arabic in vitro. The experimental data could well be described by a model in ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a134000

    authors: Kubo M,Matsuzawa Y,Yokoyama S,Tajima S,Ishikawa K,Yamamoto A,Tarui S

    更新日期:1982-09-01 00:00:00

  • A highly sensitive method for quantitative analysis of phospholipid molecular species by high-performance liquid chromatography.

    abstract::A highly sensitive method was developed for quantitative analysis of phospholipid molecular species. Diradylglycerols prepared from phospholipids with phospholipase C were converted to the anthroyl-diradylglycerol derivatives, which could be separated into molecular species and sensitively quantified by reverse-phase ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a123399

    authors: Takamura H,Kito M

    更新日期:1991-03-01 00:00:00

  • Characterization of monoclonal antibodies against human prorenin profragment and identification of active prorenins in plasma.

    abstract::Monoclonal antibodies were raised against a synthetic peptide (43 amino acid residues) that corresponds to the complete profragment of human prorenin. Seven monoclonal antibodies were chosen for further characterization. Two antibodies, 2-X-C1 and 4-X-E1, reacted with the middle region and C-terminus of the profragmen...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a122869

    authors: Ishizuka Y,Iizumi S,Akiyama F,Hori H,Poorman RA,Lobl TJ,Murakami K

    更新日期:1989-09-01 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

  • Activation by ATP of calcium-dependent NADPH-oxidase generating hydrogen peroxide in thyroid plasma membranes.

    abstract::An H2O2-generating fraction was prepared from porcine thyroid homogenate by differential and Percoll-density gradient centrifugations. The fraction consisted of mainly fragmented plasma membranes as judged by marker enzyme analysis and electron microscopy. The fraction produced H2O2 by reaction with NADPH only in the ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a122150

    authors: Nakamura Y,Ogihara S,Ohtaki S

    更新日期:1987-11-01 00:00:00

  • Conversion of pyridylamino sugar chains to corresponding reducing sugar chains.

    abstract::Pyridylamino sugar chains were converted to the corresponding reducing sugar chains by first converting them to 1-amino-1-deoxy derivatives using the method previously reported [S. Hase, J. Biochem. 112, 266-268 (1992)] and then converting the products to the corresponding reducing sugar chains using the Sommlet react...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvg112

    authors: Takahashi C,Nakakita S,Hase S

    更新日期:2003-07-01 00:00:00

  • Determination of the native subunit pattern of gizzard tropomyosin using antibodies specific to each subunit.

    abstract::The gizzard tropomyosin molecule is composed of two subunits at 1:1 molar ratio. Possible composites of the tropomyosin molecule are two kinds of homodimer (one for each subunit), a heterodimer of two subunits, or a mixture of heterodimer and homodimer(s). We tried to evaluate the native subunit composition of gizzard...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a123453

    authors: Nakamura S,Sakurai T,Nonomura Y

    更新日期:1991-05-01 00:00:00

  • Anomalous excitonic CD of meso-tetrakis(3-N-methylpyridiniumyl)porphyrin bound to poly[d(A-T)(2)].

    abstract::When meso-tetrakis(3-N-methylpyridiniumyl)porphyrin (m-TMPyP) formed a complex with poly[d(A-T)(2)], an intense bisignate excitonic CD in the Soret absorption region was observed. The excitonic CD of the m-TMPyP-poly[d(A-T)(2)] complex is unique in that no other combination of the related porphyrin, namely, meso-tetra...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvg046

    authors: Lee YA,Lee S,Lee HM,Lee CS,Kim SK

    更新日期:2003-03-01 00:00:00

  • A simple and rapid method to remove light chain phosphatase from chicken gizzard myosin.

    abstract::A simple and rapid method to remove myosin light chain phosphatase from gizzard myosin using hydroxyapatite chromatography was developed. Stable phosphorylated myosin without light chain phosphatase and kinase was also prepared by the same procedure. A convenient method using the urea gel system to detect minor contam...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a134870

    authors: Nakamura S,Nonomura Y

    更新日期:1984-08-01 00:00:00

  • Heat shock protein 90 strongly stimulates the binding of purified estrogen receptor to its responsive element.

    abstract::We previously showed that the 9 S estrogen receptor (ER) could be reconstituted from purified ER and purified heat shock protein 90 (hsp 90). So, we investigated the role of hsp 90 in the binding of purified ER to an estrogen responsive element (ERE) by using the reconstitution system. ER purified from calf uterus sho...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a124593

    authors: Inano K,Curtis SW,Korach KS,Omata S,Horigome T

    更新日期:1994-10-01 00:00:00

  • A cholesterol-independent membrane microdomain serves as a functional counter-receptor for E-selectin at the Colo201 cell surface and initiates signalling on E-selectin binding.

    abstract::The present study demonstrates that the functional counter-receptors for E-selectin at the cell surface of Colo201 human colon cancer cells are localized in detergent-insoluble membrane microdomains (DIM). Following isolation of counter-receptors from whole cell lysates using E-selectin-coupled magnetic beads followed...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvm107

    authors: Suzuki C,Kojima N

    更新日期:2007-07-01 00:00:00

  • Binding kinetics and physical properties of androgen receptor in androgen-dependent Shionogi mammary carcinoma 115.

    abstract::The characteristics of the androgen receptor in the cytoplasmic fraction of Shionogi carcinoma 115 were studied in vitro by means of charcoal adsorption assay, sucrose density gradient centrifugation, and polyacrylamide gel electrophoresis. The equilibrium dissociation constant for [3H]dihydrotestosterone (Kd = (2-5) ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a133238

    authors: Nohno T

    更新日期:1981-02-01 00:00:00

  • A new 220,000 dalton protein located in the Z lines of vertebrate skeletal muscle.

    abstract::A new protein with a chain weight of approximately 220,000 was isolated from 0.6 M KI extracts of I-Z-I brushes of rabbit and chicken skeletal muscles, using (NH4)2SO4 precipitation and three column chromatographic procedures in succession. It was only possible to separate the high molecular weight protein from actin ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a133401

    authors: Muguruma M,Kobayashi K,Fukazawa T,Ohashi K,Maruyama K

    更新日期:1981-06-01 00:00:00

  • Isolation and some properties of a new fiber-forming protein from Tetrahymena pyriformis.

    abstract::Isolation and characterization of a fibrous protein component which might be associated with contractile ring of a dividing Tetrahymena cell were attempted by making use of coprecipitation of the protein with rabbit skeletal muscle myosin. The protein was purified to homogeneity by ammonium sulfate fractionation betwe...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a133119

    authors: Numata O,Yasuda T,Hirabayashi T,Watanabe Y

    更新日期:1980-11-01 00:00:00

  • Molecular properties and functions in vitro of chicken smooth-muscle alpha-actinin in comparison with those of striated-muscle alpha-actinins.

    abstract::alpha-Actinin purified from chicken gizzard smooth muscle was characterized in comparison with alpha-actinins from chicken striated muscles, or fast-skeletal muscle, slow-skeletal muscle, and cardiac muscle. The gizzard alpha-actinin molecule consisted of two apparently identical subunits with a molecular weight of 10...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a134070

    authors: Endo T,Masaki T

    更新日期:1982-11-01 00:00:00

  • The complete amino acid sequence of coagulogen isolated from Southeast Asian horseshoe crab, Carcinoscorpius rotundicauda.

    abstract::The complete amino acid sequence of coagulogen purified from the hemocytes of the horseshoe crab Carcinoscorpius rotundicauda was determined by characterization of the NH2-terminal sequence and the peptides generated after digestion of the protein with lysyl endopeptidase, Staphylococcal aureus protease V8 and trypsin...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a135283

    authors: Srimal S,Miyata T,Kawabata S,Miyata T,Iwanaga S

    更新日期:1985-08-01 00:00:00

  • Microtubule-associated proteins, MAP 1A and MAP 1B, interact with F-actin in vitro.

    abstract::Microtubule-associated protein (MAP) 1 consisting of MAP 1A and 1B was purified from rat brain by the poly-L-aspartic acid (PLAA) method. We found that MAP 1 bound to F-actin in vitro up to a molar ratio of MAP 1 to actin monomers of 1:10. The apparent binding constant was about 2.7 x 10(7) M-1. In contrast to the bin...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a124263

    authors: Fujii T,Watanabe M,Ogoma Y,Kondo Y,Arai T

    更新日期:1993-12-01 00:00:00

  • Steroid monooxygenase of Rhodococcus rhodochrous: sequencing of the genomic DNA, and hyperexpression, purification, and characterization of the recombinant enzyme.

    abstract::Steroid monooxygenase of Rhodococcus rhodochrous is a Baeyer-Villigerase catalyzing the insertion of an oxygen atom between the C(17)- and C(20)-carbons of progesterone to produce testosterone acetate. The 5.1-kbp-long BamHI DNA fragment containing the steroid monooxygenase gene, smo, was cloned from the chromosomal D...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a022494

    authors: Morii S,Sawamoto S,Yamauchi Y,Miyamoto M,Iwami M,Itagaki E

    更新日期:1999-09-01 00:00:00

  • Structural and functional characterization of haemocyanin from the anemone hermit crab Dardanus calidus.

    abstract::Oxygen-binding to haemocyanin (Hc) is generally an exothermic process, with overall enthalphy of oxygenation varying from species to species. A number of crustacean Hcs showed a null or reduced enthalphy of oxygenation, among others, the anomuran Pagurus bernhardus and Paralithodes camtscaticae possess a completely te...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvm210

    authors: Podda G,Manconi B,Olianas A,Pellegrini M,Messana I,Mura M,Castagnola M,Giardina B,Sanna MT

    更新日期:2008-02-01 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

  • The OsGEN-L protein from Oryza sativa possesses Holliday junction resolvase activity as well as 5'-flap endonuclease activity.

    abstract::OsGEN-L has a 5'-flap endonuclease activity and plays an essential role in rice microspore development. The Class 4 RAD2/XPG family nucleases, including OsGEN-L, were recently found to have resolving activity for the Holliday junction (HJ), the intermediate of DNA strand recombination. In this study, we performed a de...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvr145

    authors: Yang Y,Ishino S,Yamagami T,Kumamaru T,Satoh H,Ishino Y

    更新日期:2012-03-01 00:00:00

  • Static and kinetic studies by fluorometry on the interaction between gluconolactone and glucoamylase from Rh. niveus.

    abstract::A transition-state analog, gluconolactone, was found to partially quench the protein fluorescence of glucoamylase [EC 3.2.1.3] from Rhizopus niveus. The interaction between gluconolactone and the enzyme was studied statically and kinetically at pH 4.5 in terms of fluorescence change. The dissociation constant Kd of th...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:

    authors: Ohnishi M,Yamashita T,Hiromi K

    更新日期:1977-01-01 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

  • Regulation of antiviral innate immune signaling by stress-induced RNA granules.

    abstract::Activation of antiviral innate immunity is triggered by cellular pattern recognition receptors. Retinoic acid inducible gene-I (RIG-I)-like receptors (RLRs) detect viral non-self RNA in cytoplasm of virus-infected cells and play a critical role in the clearance of the invaded viruses through production of antiviral cy...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章,评审

    doi:10.1093/jb/mvv122

    authors: Yoneyama M,Jogi M,Onomoto K

    更新日期:2016-03-01 00:00:00

  • Mitochondrial dynamics and interorganellar communication in the development and dysmorphism of mammalian oocytes.

    abstract::Mitochondria play many critical roles in cells, not only by supplying energy, but also by supplying metabolites, buffering Ca2+ levels and regulating apoptosis. During oocyte maturation and subsequent embryo development, mitochondria change their morphology by membrane fusion and fission, and coordinately undergo mult...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章,评审

    doi:10.1093/jb/mvz093

    authors: Udagawa O,Ishihara N

    更新日期:2020-03-01 00:00:00

  • Isolation of a 5S RNA-protein L5 complex from 60S subunits of rat liver ribosomes by cesium sulfate density-gradient equilibrium centrifugation.

    abstract::Upon CS2SO4 density-gradient equilibrium centrifugation, 60S subunits of rat liver ribosomes were dissociated to form three bands at the densities of 1.55, 1.40, and 1.30 g/ml. The bands at 1.55 and 1.30 g/ml were shown to contain exclusively RNA or proteins, respectively, whereas the band at 1.40 g/ml contained both ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a133504

    authors: Isoda N,Tanaka T,Ishikawa K

    更新日期:1981-08-01 00:00:00

  • Proteasome inhibitors block Ras/ERK signaling pathway resulting in the downregulation of Fas ligand expression during activation-induced cell death in T cells.

    abstract::Activation-induced cell death (AICD) plays a critical role in the maintenance of homeostasis and peripheral tolerance in the immune system, and is mediated by Fas ligand (FasL) expression and the interaction between Fas and FasL. In the present study, we examined the role of the ubiquitin-proteasome system in AICD usi...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a003106

    authors: Tanimoto Y,Kizaki H

    更新日期:2002-03-01 00:00:00

  • Cloning and expression of subtilisin amylosacchariticus gene.

    abstract::The gene encoding subtilisin Amylosacchariticus from Bacillus subtilis var. amylosacchariticus was isolated and the entire nucleotide sequence of the coding sequence was determined. The deduced amino acid sequence revealed an N-terminal signal peptide and pro-peptide of 106 residues followed by the mature protein comp...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a122380

    authors: Yoshimoto T,Oyama H,Honda T,Tone H,Takeshita T,Kamiyama T,Tsuru D

    更新日期:1988-06-01 00:00:00