Abstract:
:A synchronous enzyme-reaction system using water-soluble formazan and a non-enzymatic electron mediator was developed and applied to an enzyme immunoassay (EIA). The reaction system consists of four steps: (I) dephosphorylation of NADP(+) to produce NAD(+) by alkaline phosphatase (ALP), (II) reduction of NAD(+) to produce NADH with oxidation of ethanol to yield acetaldehyde by alcohol dehydrogenase (ADH), (III) reduction of water-soluble tetrazolium salt (WST-1) to produce formazan by NADH via 1-methoxy-5-methyl-phenazinium methyl sulfate (PMS), and (IV) re-reduction of NAD(+) to produce NADH by ADH. During each cycle, one molecule of tetrazolium is converted to one molecule of formazan. The concentration of formazan during the reaction was given by second-order polynomials of the reaction time. Kinetic studies strongly suggested that the synchronous enzyme-reaction system had the potential to detect an analyte at the attomole level in EIA. On the basis of the kinetic studies, optimal conditions for EIA incorporating the synchronous system were examined. NADP(+) was purified thoroughly to remove minor traces of NAD(+) in the preparation, and an ADH preparation contaminated with the lowest level of ALP activity was used. When the synchronous system was applied to a sandwich-type EIA for human C-reactive protein, the protein was detected with a sensitivity of 50 attomole per well of a micro-titer plate (0.1 ml) in a 1-h reaction. In addition, EIA with water-soluble formazan showed a more quantitative and sensitive result than that with insoluble formazan. These findings indicated that the (WST-1)-PMS system introduced in this study has a great potential for highly sensitive enzyme immunoassay.
journal_name
J Biochemjournal_title
Journal of biochemistryauthors
Inouye K,Ueno I,Yokoyama S,Sakaki Tdoi
10.1093/oxfordjournals.jbchem.a003082keywords:
subject
Has Abstractpub_date
2002-01-01 00:00:00pages
97-105issue
1eissn
0021-924Xissn
1756-2651journal_volume
131pub_type
杂志文章abstract::An endogenous inhibitor of calcium-activated neutral protease (CANP), which was isolated from rabbit skeletal muscle with chemically drastic pretreatments, comprised major (high-molecular-weight form, HMW-inhibitor) and minor (low-molecular-weight form, LMW-inhibitor) components. HMW-inhibitor was purified to homogene...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a135333
更新日期:1985-09-01 00:00:00
abstract::Stable cell lines expressing the gastric proton pump alpha- and/or beta-subunits were constructed. The cell line co-expressing the alpha- and beta-subunits showed inward Rb(+) transport, which was activated by Rb(+) in a concentration-dependent manner. In the alpha+beta-expressing cell line, rapid recovery of intracel...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a003183
更新日期:2002-06-01 00:00:00
abstract::X-ray diffraction photographs of a chicken gizzard G-actin.DNase I complex crystal have been recorded using the synchrotron radiation beam emitted by the Synchrotron Radiation Source at Daresbury and the Photon Factory at Tsukuba. The resolution limit was extended to 2.4 A and the exposure time was reduced approximate...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a134682
更新日期:1984-03-01 00:00:00
abstract::The molecular masses of two of the four DNA polymerase alpha-primase complex subunit peptides from various mammalian cells have been compared through the use of specific monoclonal antibodies. One monoclonal antibody (E4) binds to 77-kDa peptide from HeLa cells and cognate peptides from other mammalian cells (monkey, ...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a123081
更新日期:1990-04-01 00:00:00
abstract::Human liver cytosol contains multiple forms of 3 alpha-hydroxysteroid dehydrogenase and dihydrodiol dehydrogenase with hydroxysteroid dehydrogenase activity, and multiple cDNAs for the enzymes have been cloned from human liver cDNA libraries. To understand the relationship of the multiple enzyme froms to the genes, a ...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a124904
更新日期:1995-08-01 00:00:00
abstract::Type A atrial natriuretic peptide (ANP) receptor was demonstrated to be present as a tetramer in the bovine adrenal cortex. Type A ANP receptor is composed of two functional domains, namely extracellular ANP-binding and cytoplasmic guanylate cyclase domains, and generally considered to be present as a single polypepti...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a123539
更新日期:1991-07-01 00:00:00
abstract::RPE65 is a membrane-associated retinoid isomerase involved in the visual cycle responsible for sustaining vision. Many mutations in the human RPE65 gene are associated with distinct forms of retinal degenerative diseases. The pathogenic mechanisms for most of these mutations remain poorly understood. Here, we show tha...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/jb/mvv028
更新日期:2015-08-01 00:00:00
abstract::We isolated three genomic clones of rat P-450(11 beta) genes (CYP11B). Two of them corresponded to 11 beta-hydroxylase gene (CYP11B1) and aldosterone synthase gene (CYP11B2), respectively. The third one was a novel gene resembling both CYP11B1 and CYP11B2, and was named CYP11B3 gene (CYP11B3). CYP11B2 and CYP11B3 are ...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a124018
更新日期:1993-02-01 00:00:00
abstract::The vitamin A derivative, retinoic acid (RA) has various biological effects in mammalian cells and tissues. It is well known that RA induces differentiation of leukemia cells and inhibits cell growth. There are two pathways for RA action; one via RA nuclear receptors (RARs), and one via acylation of proteins by RA (re...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/jb/mvn071
更新日期:2008-09-01 00:00:00
abstract::Rat peritoneal macrophages were shown to have two distinct mannose/fucose/N-acetylglucosamine-specific lectins. The major lectin of 180 kDa, which is similar in size to the mannose receptor first isolated from alveolar macrophages (Wileman, T.E., Lennartz, M.R., & Stahl, P.D. (1986) Proc. Natl. Acad. Sci. U.S. 83, 250...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:
更新日期:1988-10-01 00:00:00
abstract::We previously reported the utilization of antisense RNA in the development of a novel immune system against RNA coliphage SP proliferation (Hirashima et al. [1986] Proc. Natl. Acad. Sci. U.S. 83, 7726-7730). We attempted to determine the most effective (i.e., those eliciting antiviral activity) sequences for targeting...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a122807
更新日期:1989-07-01 00:00:00
abstract::Tn-antigen (GalNAcα-Ser/Thr) is one of the most common aberrations associated with cancer progression and metastasis, and thus is an excellent target for development of cancer diagnostics and therapeutics. MLS128 monoclonal antibody (mAb), derived from a mouse immunized with human colon carcinoma cells, was reported t...
journal_title:Journal of biochemistry
pub_type: 杂志文章,评审
doi:10.1093/jb/mvt052
更新日期:2013-08-01 00:00:00
abstract::The most reactive single thiol group of rabbit skeletal muscle phosphofructokinase per protomer was modified with the following thiol reagents: iodoacetamide, iodoacetate, 2-hydroxyethyl disulfide, 3,3'-dithiodipropionate, and glutathione disulfide. As a result of the modification, there was increase in not only the a...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:
更新日期:1991-02-01 00:00:00
abstract::Dithiothreitol or glutathione, essential for amino acid incorporation into polypeptides in cell-free systems, inhibited glucosamine incorporation into ovalbumin, a glycoprotein containing about three residues of N-acetylucosamine and about five residues of mannose. However, the thiol was necessary for mannose incorpor...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a131155
更新日期:1976-05-01 00:00:00
abstract::The conformation and stabilities of the CL fragment isolated from a type lambda Bence Jones protein and the fragment in which the intrachain disulfide bond had been reduced were studied by measuring CD, fluorescence, and ultraviolet absorption. The results indicated that no great conformational change occurs on reduct...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a132661
更新日期:1979-11-01 00:00:00
abstract::The existence of specific binding proteins for alpha-casein, a well-known potent chemoattractant, on guinea pig peritoneal macrophages was demonstrated. Binding of 3H-alpha-casein to macrophages was found to be specific and reversible. Its association constant and the number of binding sites were 5.0 X 10(-6) M and 7....
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a132883
更新日期:1980-05-01 00:00:00
abstract::The betagamma subunits of G proteins were coimmunoprecipitated with antibodies against various alpha subunits, and analyzed by silver stain and immunoblotting with conventional transfer procedure and membrane-blocking buffer containing 2% BSA. Multiple isoforms of gamma were coimmunoprecipitated with no significant di...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a022196
更新日期:1998-11-01 00:00:00
abstract::The contribution of the P1' residue at the first reactive site of peanut protease inhibitor B-III to the inhibition was analyzed by replacement of the P1' Arg(11) with other amino acids (Arg, Ser, Ala, Leu, Phe, Asp) after selective modification of the second reactive site. The Arg derivative had the same trypsin inhi...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/jb/101.3.723
更新日期:1987-03-01 00:00:00
abstract::Several kinds of hydrophilic proteins were examined to determine their interaction with artificial liposomes. Mitochondrial aspartate aminotransferase (m-GOT) [EC 2.6.1.1], as well as cytochrome c, was found to interact strongly with negatively charged liposomes. In each case, an appreciable amount of the protein boun...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:
更新日期:1979-05-01 00:00:00
abstract::The Basidiomycete ras gene possesses a pyrimidine-rich stretch (CT-motif) with a short (7 bases) mirror repeat in which its major transcription start point is contained. To analyze the tertiary structure induced by the CT/AG-biased sequence and its effect on gene expression in supercoiled plasmids in Escherichia coli,...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a021811
更新日期:1997-10-01 00:00:00
abstract::Lambda dv plasmids having an amber mutation in an initiator gene, O or P, were constructed from mutant lambda phages by recombinant DNA techniques and several properties of such derivatives were investigated. These plasmids are perpetuated in suppressor-plus (amber-permissive) cells, but not in non-suppressor cells. T...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a134360
更新日期:1983-08-01 00:00:00
abstract::Acetohydroxy acid synthetase [EC 4.1.3.18] of Neurospora crassa has been solubilized from a mitochondrial pellet fraction by sonic treatment in 1.0 M potassium phosphate buffer at pH 7.5 containing 10 mM MgSO4 and centrifugation at 180,000 X g for 20 min. The soluble enzyme thus obtained has a high specific activity a...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:
更新日期:1975-01-01 00:00:00
abstract::The active site of thermolysin is composed of one zinc ion and five polypeptide regions [N-terminal sheet (Asn112-Trp115), alpha-helix 1 (Val139-Thr149), C-terminal loop 1 (Asp150-Gly162), alpha-helix 2 (Ala163-Val176) and C-terminal loop 2 (Gln225-Ser234)]. To explore their catalytic roles, we introduced single amino...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/jb/mvn140
更新日期:2009-01-01 00:00:00
abstract::Manganese ion, like Mg2+, has been found to produce high biosynthetic activity of the unadenylylated form of glutamine synthetase obtained from Mycobacterium smegmatis, and the activity with each of these cations was decreased by the adenylylation of the enzyme. Further, the gamma-glutamyltransferase reaction was cata...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a135145
更新日期:1985-04-01 00:00:00
abstract::MicroRNAs (miRNAs) are a class of ∼22 nt non-coding RNAs that control diverse biological functions in animals, plants and unicellular eukaryotes by promoting degradation or inhibition of translation of target mRNAs. miRNA expression is often tissue specific and developmentally regulated. Aberrant expression of miRNAs ...
journal_title:Journal of biochemistry
pub_type: 杂志文章,评审
doi:10.1093/jb/mvq096
更新日期:2010-10-01 00:00:00
abstract::Highly purified recombinant human tumor necrosis factor (TNF) (molecular mass determined as 17 kilodaltons (kDa) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and as 36 kDa by Sephadex G-100 gel chromatography) was labeled with 125I to a specific activity of 5 microCi/micrograms without appreciable loss...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a121744
更新日期:1986-09-01 00:00:00
abstract::Deoxyhypusine synthase catalyzes the NAD+-dependent formation of deoxyhypusine in the eIF-5A precursor protein by transferring the 4-aminobutyl moiety of spermidine. This enzyme has recently been shown to be essential for cell viability and growth of yeast [Sasaki, K., Abid, M.R., and Miyazaki, M. (1996) FEBS Lett. 38...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a021652
更新日期:1997-04-01 00:00:00
abstract::The chromatin remodeling complex, SWI/SNF, is known to regulate the transcription of several genes by altering the chromatin structure in an ATP-dependent manner. SWI/SNF exclusively contains BRG1 or BRM as an ATPase subunit. In the present study, we studied the role of SWI/SNF containing BRM or BRG1 in the expression...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/jb/mvi147
更新日期:2005-10-01 00:00:00
abstract::Heptic fibrosis/cirrhosis is a common hepatic disease characterized by the hyper-accumulation of connective tissue components, and hepatic necrosis. Chronic alcohol ingestion, viral infection, and metabolic disorders are contributing factors and there has been no effective treatment. Hepatocyte growth factor (HGF), or...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a124958
更新日期:1995-09-01 00:00:00
abstract::Mild trypsinization of human erythrocyte stroma solubilized CR1 (complement receptor type 1, C3b/C4b receptor) without significant loss of decay-accelerating activity to C5 convertases on hemolytic intermediate cells (EAC 1-3b, P). The solubilized CR1 was purified using DEAE-Sephacel, C3-Sepharose, and anti-CR1-Sephar...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a121823
更新日期:1986-11-01 00:00:00