Stabilization of human RNase 1 by introduction of a disulfide bond between residues 4 and 118.

Abstract:

:In order to stabilize human RNase 1 by introduction of an intramolecular cross-link, a mutant protein (4-118CL RNase 1), in which Arg4 and Val118 are replaced with cysteine residues and linked by a disulfide bond, was designed and expressed in Escherichia coli as inclusion bodies. The 4-118CL RNase 1 that refolded under redox conditions was a monomer without free SH groups and retained 11% of the activity of the wild-type recombinant RNase 1, indicating that the mutant enzyme was correctly folded with the formation of an additional disulfide bond between Cys4 and Cys118. From guanidium chloride denaturation experiments based on the assumption of a two-state transition for unfolding, it was demonstrated that the introduction of the present cross-link increased the thermodynamic stability of RNase 1 by 2.0 kcal/mol. This value was lower than that, 5.4 kcal/mol, theoretically calculated from the reduction of chain entropy of the unfolded state due to the introduction of the cross-link. These results suggest that the present cross-link also destabilized the folded state of RNase 1 by 3.4 kcal/mol. Along with the increase in the thermodynamic stability, the stability of RNase 1 against trypsin digestion was also significantly increased by the introduction of this cross-link. It is likely, although not proven, that stabilized human RNases are favorable for clinical use, because human RNase-based immunotoxins should have long half-lives as to proteolytic degradation after endocytosis.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Futami J,Tada H,Seno M,Ishikami S,Yamada H

doi

10.1093/oxfordjournals.jbchem.a022747

keywords:

subject

Has Abstract

pub_date

2000-08-01 00:00:00

pages

245-50

issue

2

eissn

0021-924X

issn

1756-2651

journal_volume

128

pub_type

杂志文章
  • 5'-N-ethylcarboxamido [3H] adenosine binding sites of mouse P815 mastocytoma cell membranes: solubilization and partial purification by affinity chromatography.

    abstract::A 5'-N-ethylcarboxamido[3H]adenosine ([3H]NECA) binding site of mouse mastocytoma P815 cell membranes has been purified approximately 100-fold by affinity chromatography. This adenosine binding site, which has a similar specificity to that of the A2 adenosine receptor, was absorbed on NECA-linked Sepharose 6B and elut...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:

    authors: Nakata H

    更新日期:1989-05-01 00:00:00

  • Biosynthesis of rat kidney catalase. Double-labeling by [14C]leucine and delta-amino[3H]levulinic acid.

    abstract::Kidney catalase [EC 1.11.1.6] was doubly labeled by injecting a mixture of [14C]-leucine and delta-amino[3H]levulinic acid (ALA) into normal and allyliso-(2-isopropyl-4-pentenoyl)urea (Sedormid)-treated rats. Shortly after the injection of a tracer dose or the precursors, catalase in microsomes showed the highest spec...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a131272

    authors: Higashi T,Kawamata F,Dogen M

    更新日期:1976-08-01 00:00:00

  • Binding strength of calcium ion to bovine alpha-lactalbumin.

    abstract::A Ca2+-sensitive electrode was used for determination of the binding strength of Ca2+ to bovine alpha-lactalbumin in 60 mM Tris buffer (pH 7.8-8.5) in the presence of various concentrations of NaCl. The dependence of the apparent binding constant on the concentration of NaCl was consistent with competitive binding of ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a121869

    authors: Hamano M,Nitta K,Kuwajima K,Sugai S

    更新日期:1986-12-01 00:00:00

  • gamma-Carboxyglutamic acid (Gla)-domainless blood coagulation factor IXa species: preparation and properties.

    abstract::To investigate the function of the gamma-carboxyglutamic acid (Gla) residues of factor IXa in the activation of factor X, a new species of bovine factor IXa, designated "factor IXa beta'," and its corresponding Gla-domainless form, designated "Gla-domainless factor IXa beta'," were prepared under controlled conditions...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a123701

    authors: Morita T,Fukudome K,Miyata T,Iwanaga S

    更新日期:1991-12-01 00:00:00

  • Permeability change of liposomal membrane induced by interleukin-1 alpha.

    abstract::Interleukin-1 (IL-1) is produced and released by various cells, including activated macrophages, and plays important roles in inflammation as well as immune responses. Since the precursor of IL-1 has no signal peptide, the mechanism of IL-1 release has been an enigma. To investigate the possibility of direct interacti...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a124987

    authors: Oku N,Saito N,Okada S,Watanabe N,Kobayashi Y

    更新日期:1995-10-01 00:00:00

  • Comparative studies of protein properties and bacteriochlorophyll contents of bacteriochlorophyll-protein complexes from spectrally different types of Rhodopseudomonas palustris.

    abstract::The amino acid compositions, constituent polypeptides and bacteriochlorophyll (Bchl) contents of two kinds of Bchl-protein complexes isolated from Rhodopseudomonas palustris were examined. Spectrally dissimilar intracytoplasmic membranes obtained from cells cultured under different conditions were used as starting mat...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a134110

    authors: Hayashi H,Nakano M,Morita S

    更新日期:1982-12-01 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

  • Immunochemical studies on non-precipitating guinea pig antibody produced by administration of an excessive dose of Bacillus subtilis alpha-amylase.

    abstract::Administration of an excessive dose of Bacillus subtilis alpha-amylase [EC 3.2.1.1, alpha-1,4-glucan 4-glucanohydrolase] (BalphaA) induced the production of non-precipitating (non-ppt) IgG2 antibody in guinea pigs, whereas immunization with a normal dose produced precipitating (ppt) IgG1 and IgG2 antibodies. The non-p...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a131657

    authors: Mori N,Nakamura T,Koyama J

    更新日期:1977-07-01 00:00:00

  • Chemical modification of the Ca2+-dependent ATPase of sarcoplasmic reticulum from skeletal muscle. I. Binding of N-ethylmaleimide to sarcoplasmic reticulum: evidence for sulfhydryl groups in the active site of ATPase and for conformational changes induced

    abstract::The time course of binding of N-ethylmaleimide (NEM) to the SR was measured at pH 7.5 in the presence or absence of ATP or ADP. The following results were obtained. 1. Both in the presence and absence of nucleotide, the ATPase [EC 3.6.1.3] activity decreased linearly with increase in the amount of NEM bound to the fra...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a131109

    authors: Yoshida H,Tonomura Y

    更新日期:1976-03-01 00:00:00

  • Studies on the physicochemical structure and stability of an active fragment (T2A) of colicin E3.

    abstract::The physicochemical nature of an active fragment (T2A) of colicin E3 was examined together with those of its specific inhibitor, B, and the T2A-B complex under various conditions by means of CD and fluorescence spectroscopy. 1. T2A was mainly composed of unordered structure, although the molecule was folded into a rat...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a132805

    authors: Suzuki K,Ohno S,Imahori K

    更新日期:1980-03-01 00:00:00

  • Interaction between pullulanase from Klebsiella pneumoniae and cyclodextrins.

    abstract::The interaction between pullulanase from Klebsiella pneumoniae and alpha-, beta-, and gamma-cyclodextrins and 6-O-alpha-glucosyl-alpha-cyclodextrin and 6-O-alpha-glucosyl-beta-cyclodextrin was examined by means of inhibition studies of the enzyme activity, UV difference spectroscopy, and flow calorimetry. All the abov...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a124010

    authors: Iwamoto H,Ohmori M,Ohno M,Hirose J,Hiromi K,Fukada H,Takahashi K,Hashimoto H,Sakai S

    更新日期:1993-01-01 00:00:00

  • Inversions in mitochondrial DNA of a petite mutant of yeast.

    abstract::Mitochondrial DNA of an erythromycin-resistant petite mutant of yeast, E734, showed physical maps of inversion, which occurred between two cross-over sites in the fragments Hae A and Hae B. The pair of cross-over sites was inferred to be accommodated within a repeat unit of E734 mtDNA during petite mutation by joining...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a134985

    authors: Wakabayashi K,Furutani Y

    更新日期:1984-11-01 00:00:00

  • Nucleotide sequence of the gene encoding NADH dehydrogenase from an alkalophile, Bacillus sp. strain YN-1.

    abstract::The gene encoding NADH dehydrogenase from an alkalophile, Bacillus sp., was cloned and sequenced. The cloned DNA fragment contained an open reading frame of 1,557 nucleotides which encodes a polypeptide composed of 519 amino acid residues (Mr 55,830). The predicted amino acid sequence was consistent with the partial a...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a123440

    authors: Xu XM,Koyama N,Cui M,Yamagishi A,Nosoh Y,Oshima T

    更新日期:1991-05-01 00:00:00

  • N- and C-terminal fragments of a globular protein constructed by elongation of modules as a units associated for functional complementation.

    abstract::We have been interested in partially folded proteins with marginal stability and activity, because they have a potential to be mature proteins by artificial evolution. A module is defined as a contiguous peptide chain forming a compact region in a globular protein. Modules may be used as building blocks to create part...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvn099

    authors: Tsuji T,Nagata T,Yanagawa H

    更新日期:2008-10-01 00:00:00

  • Identification of the catalytic residues of sequence-specific and histidine-free ribonuclease colicin E5.

    abstract::Colicin E5 cleaves tRNAs for Tyr, His, Asn and Asp in their anticodons to abolish protein synthesis in Escherichia coli. We previously showed how its C-terminal RNase domain, E5-CRD, recognizes the anticodon bases but the catalytic mechanism remained to be elucidated. Although the reaction products with 5'-OH and 2',3...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvs077

    authors: Inoue-Ito S,Yajima S,Fushinobu S,Nakamura S,Ogawa T,Hidaka M,Masaki H

    更新日期:2012-10-01 00:00:00

  • Purification of dihydrofolate reductase amplified in Escherichia coli K-12.

    abstract::The Escherichia coli strain carrying pTP 6-10 which was constructed in our previous work (Iwakura, M., et al. (1983) J. Biochem. 93, 927-930) produces more than 400-fold dihydrofolate reductase as compared with the strain without the plasmid. Dihydrofolate reductase was highly purified from the cell-free extract of th...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a134400

    authors: Iwakura M,Shimura Y,Sakai T,Tsuda K

    更新日期:1983-09-01 00:00:00

  • A 13C-NMR study on the interaction of riboflavin with egg white riboflavin binding protein.

    abstract::The interaction between riboflavin and riboflavin binding protein (RBP) was studied by 13C-NMR spectroscopy. The 13C-NMR spectra of riboflavin selectively enriched at the 2-, 4-, 4a-, and 10a-positions and of (3-[13C]methyl)riboflavin were measured both in the free and RBP-bound forms. The 13C signals of 13C-enriched ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a134813

    authors: Miura R,Tojo H,Fujii S,Yamano T,Miyake Y

    更新日期:1984-07-01 00:00:00

  • Alpha-2-macroglobulin-like protease inhibitor from the egg white of cuban crocodile (Crocodylus rhombifer).

    abstract::A high molecular weight protease inhibitor was purified from the egg white of Cuban crocodile (Crocodylus rhombifer). It inhibited the casein hydrolyzing activity of trypsin, subtilisin and papain. Its native molecular weight was 730,000 and it consisted of four subunits of equal molecular weight, each pair of which w...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a134145

    authors: Ikai A,Kitamoto T,Nishigai M

    更新日期:1983-01-01 00:00:00

  • Interaction of the Pseudomonas cepacia DSM3959 lipase with its chaperone, LimA.

    abstract::The lipA gene of Pseudomonas cepacia DSM3959 requires a downstream gene, limA, in oder to express lipase activity. The product of the lim gene, LimA, is a molecular chaperone required during the folding of lipase in oder for the lipase to adopt an active conformation. The lipase and LimA proteins have been shown to fo...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a124948

    authors: Hobson AH,Buckley CM,Jørgensen ST,Diderichsen B,McConnell DJ

    更新日期:1995-09-01 00:00:00

  • Identification of amino acid residues responsible for the changes of absorption and fluorescence spectra on the binding of subtilisin BPN' and Streptomyces subtilisin inhibitor.

    abstract::An ultraviolet absorption difference spectrum characteristic of the ionization change of a tyrosyl residue was observed on the binding of subtilisin BPN' with Streptomyces subtilisin inhibitor (SSI) at alkaline pH. This difference spectrum was considered to be induced by a pKa shift (from 9.7 to > or = 11.5) of a tyro...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a124275

    authors: Masuda-Momma K,Shimakawa T,Inouye K,Hiromi K,Kojima S,Kumagai I,Miura K,Tonomura B

    更新日期:1993-12-01 00:00:00

  • Significance of Thr182 in the nucleotide-exchange and GTP-hydrolysis reactions of the alpha subunit of GTP-binding protein Gi2.

    abstract::The crystal structures of the GTP- and GDP-bound alpha subunits of heterotrimeric GTP-binding proteins were recently determined, and a conserved Thr residue in the G2 (linker 2) region of the alpha subunits, which corresponds to Thr182 in Gi2 alpha, was deduced to interact with the gamma-phosphate of GTP and Mg2+. To ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/118.5.1083

    authors: Nishina H,Nimota K,Kukimoto I,Maehama T,Takahashi K,Hoshino S,Kanaho Y,Katada T

    更新日期:1995-11-01 00:00:00

  • Properties of proteins produced after damage to deoxyribonucleic acid of Escherichia coli.

    abstract::Large amounts of extra proteins, X (in the envelope fraction) and X' (in the cytoplasmic fraction) were detected by SDS-polyacrylamide gel electrophoresis when DNA of Escherichia coli was damaged. These two proteins had the same apparent molecular weight (appproximately 40,000) and were produced under identical condit...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:

    authors: Yoda K,Nagai K,Tamura G

    更新日期:1977-05-01 00:00:00

  • Competing solvent effects of polyols and guanidine hydrochloride on protein stability.

    abstract::The denaturation of lysozyme and ribonuclease A by guanidine hydrochloride was followed in the presence and absence of glycerol and sorbitol by means of circular dichroism measurements at 25 degrees C. The protein-solvent interactions in the presence of these polyols were also studied by means of density measurements,...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a123088

    authors: Gekko K,Ito H

    更新日期:1990-04-01 00:00:00

  • Properties of phospholipase A1/transacylase in the white muscle of bonito Euthynnus pelamis (Linnaeus).

    abstract::The properties of phospholipase A1 (PLA1) obtained from the white muscle of bonito, Euthynnus pelamis (Linnaeus), were examined. The PLA1 activity had a pH optimum from 6.5 to 7.0 for phosphatidylcholine (PC), and calcium ion was not required. The optimum temperature was from 20 to 30 degrees C. When a fatty alcohol w...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a021876

    authors: Hirano K,Tanaka A,Yoshizumi K,Tanaka T,Satouchi K

    更新日期:1997-12-01 00:00:00

  • Interaction between lipopolysaccharide and intracellular serine protease zymogen, factor C, from horseshoe crab (Tachypleus tridentatus) hemocytes.

    abstract::The interaction between lipopolysaccharide (LPS) and an LPS-sensitive serine protease zymogen, factor C, purified from horseshoe crab (Tachypleus tridentatus) hemocytes, was investigated to elucidate the LPS-mediated activation of factor C. The rate of activation of the zymogen factor C was highly dependent on the con...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a122276

    authors: Nakamura T,Tokunaga F,Morita T,Iwanaga S

    更新日期:1988-02-01 00:00:00

  • Purification and characterization of a poly(A) polymerase from beef liver nuclei.

    abstract::Poly(A) polymerase [EC 2.7.7.19] was highly purified from beef liver nuclei by the use of column chromatographies on heparin-Sepharose 4B and Blue Dextran-Sepharose 4B. The purified enzyme showed one major protein band of the molecular weight of 57,000 in SDS polyacrylamide gel electrophoresis, which agreed with the m...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a132979

    authors: Ohyama Y,Fukami H,Ohta T

    更新日期:1980-08-01 00:00:00

  • Remarkable improvement in the heat stability of CutA1 from Escherichia coli by rational protein design.

    abstract::To enhance the heat stability of the CutA1 protein from Escherichia coli (EcCutA1) so that it has comparable stability to CutA1 from Pyrococcus horikoshii with a denaturation temperature (T(d)) of 150°C, we used the Stability Profile of Mutant Protein (SPMP) to examine the structure-sequence (3D-1D) compatibility betw...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/jb/mvq079

    authors: Matsuura Y,Ota M,Tanaka T,Takehira M,Ogasahara K,Bagautdinov B,Kunishima N,Yutani K

    更新日期:2010-10-01 00:00:00

  • Oxygen metabolism of human colostral macrophages: comparison with monocytes and polymorphonuclear leukocytes.

    abstract::Human colostral macrophages stimulated by opsonized zymosan or phorbol myristate acetate (PMA) released superoxide anions (O2-) and hydrogen peroxide (H2O2) with activities comparable to those of monocytes and about one-fourth of those of polymorphonuclear leukocytes (PMNL) of blood. The O2- -forming oxidase in the ma...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a134727

    authors: Tsuda H,Takeshige K,Shibata Y,Minakami S

    更新日期:1984-05-01 00:00:00

  • The modification of sulfhydryl groups of glutamine synthetase from Bacillus stearothermophilus with 5, 5'-dithiobis(2-nitrobenzoic acid).

    abstract::The SH groups of glutamine synthetase [EC 6.3.1.2] from Bacillus stearothermophilus were modified with 5, 5'-dithiobis(2-nitrobenzoic acid) in order to determine the number of SH groups in the molecule as well as the effect of the modification on the enzyme activity. Three SH groups per subunit were detected after co...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a131021

    authors: Hachimori A,Takeda A,Nagaoka T,Suzuki H,Nosoh Y,Samejima T

    更新日期:1975-12-01 00:00:00

  • Inhibition of cathepsin E by alpha 2-macroglobulin and the resulting structural changes in the inhibitor.

    abstract::The activity of cathepsin E (M(r) approximately 80K), a dimeric aspartic proteinase with two active sites per molecule, toward a protein substrate (reduced and carboxymethylated ribonuclease A) was shown to be completely inhibited by alpha 2-macroglobulin (alpha 2M) at pH 5.5. On the other hand, the activity toward a ...

    journal_title:Journal of biochemistry

    pub_type: 杂志文章

    doi:10.1093/oxfordjournals.jbchem.a124077

    authors: Athauda SB,Arakawa H,Nishigai M,Takahashi T,Ikai A,Takahashi K

    更新日期:1993-05-01 00:00:00