Fast solid support detection of PCR amplified viral DNA sequences using radioiodinated or hapten labelled primers.
AuthorsSauvaigo S, Fouqué B, Roget A, Livache T, Bazin H, Chypre C, Téoule R
JournalNucleic Acids Res
PubMed ID2162518
'Oligonucleotides with novel modifications have been synthesized and incorporated into enzymatically amplified DNA sequences. They allow the fast detection of viral DNA sequences after two rounds of amplification. The hybrids formed are immobilized by affinity on coated tubes and detected by direct beta (32P) or gamma (125I) counting or by ... More
Determinants of substrate recognition in the protein-tyrosine phosphatase, PTP1.
'Photoaffinity labeling has been used to identify amino acids involved in recognition of protein substrates by the protein-tyrosine phosphatase PTP1. The photoactive amino acid p-benzoylphenylalanine (Bpa) was incorporated into a phosphotyrosine-containing peptide derived from epidermal growth factor autophosphorylation site Tyr992 (EGFR988 998). This peptide photoinactivated PTP1 in a time- and ... More
Activation of p70 S6 kinase and erk-encoded mitogen-activated protein kinases is resistant to high cyclic nucleotide levels in Swiss 3T3 fibroblasts.
'Treatment of Swiss mouse 3T3 fibroblasts with certain cyclic nucleotide phosphodiesterase inhibitors (theophylline, SQ 20,006, and MY-5445) prevents the activation of the M(r) 70,000 S6 kinase (p70S6k) induced by a variety of external stimuli. Concentrations giving half-maximal inhibition were 800, 50, and 25 microM, respectively. Western blot analysis and immunocomplex ... More
Hamycin inhibits IL-8-induced biologic response by modulating its receptor in human polymorphonuclear neutrophils.
AuthorsManna SK, Samanta S, Samanta AK
JournalJ Immunol
PubMed ID9366432
'IL-8, a neutrophil chemotactic agent, is involved in a large number of neutrophil-driven acute and chronic inflammatory diseases. We have found that hamycin, an antifungal agent, reduces IL-8-induced migration and binding of 125I-labeled IL-8 to neutrophils by 66 and 75%, respectively. Other IL-8-induced biologic functions, such as superoxide generation, intracellular ... More
Unfolding pathway in red kidney bean acid phosphatase is dependent on ligand binding.
AuthorsCashikar AG, Rao NM
JournalJ Biol Chem
PubMed ID8617740
'Structural basis for ligand-induced protein stabilization was investigated in the case of an acid phosphatase (red kidney bean purple acid phosphatase (KBPAP)) from red kidney bean. Phosphate, a physiological ligand, increases the stability against solvent denaturation by 3.5 kcal/mol. Generality of phosphate stabilization was shown by similar effects with other ... More
A comparison of the sensitivity and specificity of enzyme immunoassays and time-resolved fluoroimmunoassay.
AuthorsRoberts IM, Jones SL, Premier RR, Cox JC
JournalJ Immunol Methods
PubMed ID1919036
'Time-resolved fluoroimmunoassay (TR-FIA) and various enzyme immunoassays (EIA) were compared in order to determine the detection system which showed the greatest degree of sensitivity without sacrificing specificity. The system chosen for the evaluation of these assays was the detection of antibodies to human immunodeficiency virus (HIV). For EIA, horseradish peroxidase ... More
H2O2-induced uncoupling of bovine lens Na+,K+-ATPase.
AuthorsGarner WH, Garner MH, Spector A
JournalProc Natl Acad Sci U S A
PubMed ID6300890
'A 1-hr exposure of bovine lenses in organ culture to H2O2 concentrations in the range found in the aqueous fluid of patients with cataracts inhibits 86Rb+ influx. At 1 mM H2O2, complete inhibition was observed and further investigated. Membrane permeability is slightly decreased. Although lactate concentrations increase 2-fold, lens ATP ... More
Thioredoxin is required for vacuole inheritance in Saccharomyces cerevisiae.
AuthorsXu Z, Wickner W
JournalJ Cell Biol
PubMed ID8603912
'The vacuole of Saccharomyces cerevisiae projects a stream of tubules a and vesicles (a "segregation structure") into the bud in early S phase. We have described an in vitro reaction, requiring physiological temperature, ATP, and cytosol, in which isolated vacuoles form segregation structures and fuse. This in vitro reaction is ... More
Enzyme-linked immunosorbent assay (ELISA).
AuthorsReen DJ
JournalMethods Mol Biol
PubMed ID7951745
Alkaline phosphatase isozymes as possible markers of differentiation in human testicular teratocarcinoma cell lines.
AuthorsBenham FJ, Andrews PW, Knowles BB, Bronson DL, Harris H
JournalDev Biol
PubMed ID7308577
Secreted placental alkaline phosphatase as a eukaryotic reporter gene.
AuthorsCullen BR, Malim MH
JournalMethods Enzymol
PubMed ID1479908
Enzyme immunoassay ELISA and EMIT.
AuthorsEngvall E
JournalMethods Enzymol
PubMed ID6999296
Binding of 2'(3')-O-(2,4-6-trinitrophenyl) ADP to soluble alpha beta protomers of Na, K-ATPase modified with fluorescein isothiocyanate. Evidence for two distinct nucleotide sites.
AuthorsWard DG, Cavieres JD
JournalJ Biol Chem
PubMed ID8647832
The overall reaction of well-defined solubilized protomers of Na,K-ATPase (one alpha plus one beta subunit) retains the dual ATP dependence observed with the membrane-bound enzyme, with distinctive ATP effects in the submicromolar and submillimolar ranges (Ward, D. G., and Cavieres, J. D. (1993) Proc. Natl. Acad. Sci. U. S. A. ... More
A sensitive competitive ELISA for 2,4-dinitrophenol using 3,6-fluorescein diphosphate as a fluorogenic substrate.
AuthorsHuang Z, Olson NA, You W, Haugland RP
JournalJ Immunol Methods
PubMed ID1301064
A competitive ELISA for sensitive detection of 2,4-dinitrophenol (DNP) was established. Certain amounts of bovine serum albumin conjugate of DNP were readily coated on a polystyrene microplate. Free DNP was then quantitated by its competition with the coated DNP for binding to anti-DNP (antibody)-alkaline phosphatase conjugate. The enzyme conjugate remaining ... More
Nucleic acid detection using non-radioactive labelling methods.
AuthorsMansfield ES, Worley JM, McKenzie SE, Surrey S, Rappaport E, Fortina P
JournalMol Cell Probes
PubMed ID7477006
Nucleic acid probe-based assays are now widely used in genetic research, human identification, forensics and in a broad spectrum of clinical assays in the fields of microbiology, haematology/oncology and virology. Labelled probes are used in a variety of assay formats including dot-blots, Southern blots (DNA target), Northern blots (RNA target), ... More
Plasmalopsychosine of human brain mimics the effect of nerve growth factor by activating its receptor kinase and mitogen-activated protein kinase in PC12 cells. Induction of neurite outgrowth and prevention of apoptosis.
AuthorsSakakura C, Igarashi Y, Anand JK, Sadozai KK, Hakomori S
JournalJ Biol Chem
PubMed ID8557709
Plasmalopsychosine, a characteristic fatty aldehyde conjugate of beta-galactosylsphingosine (psychosine) found in brain white matter, enhances p140trk (Trk A) phosphorylation and mitogen-activated protein kinase (MAPK) activity and as a consequence induces neurite outgrowth in PC12 cells. The effect of plasmalopsychosine on neurite outgrowth and its prolonged activation of MAPK was similar ... More
Interaction of fluorescein isothiocyanate with the (H+ + K+)-ATPase.
AuthorsJackson RJ, Mendlein J, Sachs G
JournalBiochim Biophys Acta
PubMed ID6303415
Fluorescein isothiocyanate was used to covalently label the gastric (H+ + K+)-ATPase. FITC treatment of the enzyme inhibited the ATPase activity while largely sparing partial reactions such as the associated p-nitrophenylphosphatase activity. ATP protected against inhibition suggesting the ligand binds at or near an ATP binding site. At 100% inhibition ... More
Serum and host liver activities of glycosidases and sialyltransferases in animals bearing transplantable tumors.
AuthorsBosmann HB, Spataro AC, Myers MW
JournalRes Commun Chem Pathol Pharmacol
PubMed ID172998
Rats bearing Reuber H-35 or Novikoff hepatomas and mice bearing L1210 or L5178Y murine leukemias exhibited elevated serum levels of fetuin : N-acetylneuraminic acid transferase (EC 2.4.99.1) activity. The serum transferase activity could be correlated with the growth rate of the tumor; in animals bearing the more rapidly growing Novikoff ... More
Vascular endothelial growth factor (VEGF)-driven actin-based motility is mediated by VEGFR2 and requires concerted activation of stress-activated protein kinase 2 (SAPK2/p38) and geldanamycin-sensitive phosphorylation of focal adhesion kinase.
In endothelial cells, vascular endothelial growth factor (VEGF) induces an accumulation of stress fibers associated with new actin polymerization and rapid formation of focal adhesions at the ventral surface of the cells. This cytoskeletal reorganization results in an intense motogenic activity. Using porcine endothelial cells expressing one or the other ... More
p-nitrophenylphosphatase activity of plasma membrane H(+)-ATPase from yeast. Implications for the regulation of the catalytic cycle by H+.
AuthorsFerreira-Pereira A, Alves-Ferreira M, de Carvalho-Alves PC
JournalJ Biol Chem
PubMed ID8163511
The H(+)-ATPase from Schizosaccharomyces pombe belongs to the group of transport ATPases which displays two main conformational states, E1 and E2 (P-type ATPase). In this report, we show that, as in the case of other P-type ATPase, the purified enzyme exhibits a p-nitrophenylphosphatase activity which can be completely inhibited by ... More
Regulation of acidification and apoptosis by SHP-1 and Bcl-2.
Recruitment of the SH2 domain containing cytoplasmic protein-tyrosine phosphatase SHP-1 to the membrane by somatostatin (SST) is an early event in its antiproliferative signaling that induces intracellular acidification-dependent apoptosis in breast cancer cells. Fas ligation also induces acidification-dependent apoptosis in a manner requiring the presence of SHP-1 at the membrane. ... More
Ability of RapID Yeast Plus System to identify 304 clinically significant yeasts within 5 hours.
AuthorsKitch TT, Jacobs MR, McGinnis MR, Appelbaum PC
JournalJ Clin Microbiol
PubMed ID8727877
The RapID Yeast Plus System (Innovative Diagnostic Systems, Norcross, Ga.) is a qualitative micromethod that uses conventional and chromogenic substrates for the identification of medically important yeasts. The ability of the RapID Yeast Plus system to accurately identify 304 clinical yeast isolates within 5 h was evaluated. The RapID Yeast ... More
Effect of C2-associated carbohydrate structure on Ig effector function: studies with chimeric mouse-human IgG1 antibodies in glycosylation mutants of Chinese hamster ovary cells.
AuthorsWright A, Morrison SL
JournalJ Immunol
PubMed ID9531299
The complex biantennary oligosaccharide at Asn297 of IgG is essential for some effector functions. To investigate the effect of carbohydrate structure on Ab function, we have now expressed mouse-human chimeric IgG1 Abs in Chinese hamster ovary (CHO) cells with defined defects in carbohydrate biosynthesis. We had previously shown that IgG1 ... More
Inhibitory effect of okadaic acid on the p-nitrophenyl phosphate phosphatase activity of protein phosphatases.
AuthorsTakai A, Mieskes G
JournalBiochem J
PubMed ID1850239
The phosphatase activities of type 2A, type 1 and type 2C protein phosphatase preparations were measured against p-nitrophenyl phosphate (pNPP), a commonly used substrate for alkaline phosphatases. Of the three types of phosphatase examined, the type 2A phosphatase exhibited an especially high pNPP phosphatase activity (119 +/- 8 mumol/min per ... More
Substrate sites of the (Na+ + K+)-ATPase: pertinence of the adenine and fluorescein binding sites.
AuthorsDavis RL, Robinson JD
JournalBiochim Biophys Acta
PubMed ID2829969
The (Na+ + K+)-activated ATPase catalyzes the K+-activated hydrolysis of 3-O-methylfluorescein phosphate (3OMFP) with a Km of 50 microM, nearly two orders of magnitude lower than the Km for nitrophenyl phosphate, 3 mM. Both ATP and nitrophenyl phosphate are competitors toward 3OMFP with Ki values corresponding to their Km values ... More