Institution
Applied Biosystems
About: Applied Biosystems is a based out in . It is known for research contribution in the topics: Mass spectrometry & Capillary electrophoresis. The organization has 1521 authors who have published 1579 publications receiving 285423 citations.
Topics: Mass spectrometry, Capillary electrophoresis, Nucleic acid, Oligonucleotide, Tandem mass spectrometry
Papers published on a yearly basis
Papers
More filters
••
TL;DR: A novel cell-based functional assay to directly monitor G protein-coupled receptor (GPCR) activation in a high-throughput format, based on a common GPCR regulation mechanism, the interaction between β-arrestin and ligand-activated GPCRs, is described.
Abstract: A novel cell-based functional assay to directly monitor G protein-coupled receptor (GPCR) activation in a high-throughput format, based on a common GPCR regulation mechanism, the interaction between beta-arrestin and ligand-activated GPCR, is described. A protein-protein interaction technology, the InteraX trade mark system, uses a pair of inactive beta-galactosidase (beta-gal) deletion mutants as fusion partners to the protein targets of interest. To monitor GPCR activation, stable cell lines expressing both GPCR- and beta-arrestin-beta-gal fusion proteins are generated. Following ligand stimulation, beta-arrestin binds to the activated GPCR, and this interaction drives functional complementation of the beta-gal mutant fragments. GPCR activation is measured directly by quantitating restored beta-gal activity. The authors have validated this assay system with two functionally divergent GPCRs: the beta2-adrenergic amine receptor and the CXCR2 chemokine-binding receptor. Both receptors are activated or blocked with known agonists and antagonists in a dose-dependent manner. The beta2-adrenergic receptor cell line was screened with the LOPAC trade mark compound library to identify both agonists and antagonists, validating this system for high-throughput screening performance in a 96-well microplate format. Hit specificity was confirmed by quantitating the level of cAMP. This assay system has also been performed in a high-density (384-well) microplate format. This system provides a specific, sensitive, and robust methodology for studying and screening GPCR-mediated signaling pathways.
65 citations
••
TL;DR: A new NMR method is described for the generation of absorption mode two‐dimensional NOE spectra of oligonucleotides in H2O solution that yields spectra that are free of baseline distortions with excellent suppression of the intense H 2O resonance.
65 citations
••
TL;DR: The TaqMan‐ASA method was applied to detect a prevalent 727G>T mutation in Japanese patients with glycogen storage disease type Ia and a common 985A>G mutation in Caucasian patients with medium‐chain acyl‐CoA dehydrogenase deficiency.
Abstract: We have devised an allele-specific amplification method with a TaqMan fluorogenic probe (TaqMan-ASA) for the detection of point mutations. Pairwise PCR amplification using two sets of allele-specific primers in the presence of a TaqMan probe was monitored in real time with a fluorescence detector. Difference in amplification efficiency between the two PCR reactions was determined by "threshold" cycles to differentiate mutant and normal alleles without post-PCR processing. The method measured the efficiency of amplification rather than the presence or absence of end-point PCR products, therefore allowing greater flexibility in designing allele-specific primers and an ample technical margin for allelic discrimination. We applied the TaqMan-ASA method to detect a prevalent 727G>T mutation in Japanese patients with glycogen storage disease type Ia and a common 985A>G mutation in Caucasian patients with medium-chain acyl-CoA dehydrogenase deficiency. The method can be automated and may be applicable to the DNA diagnosis of various genetic diseases.
65 citations
••
TL;DR: Analysis of synthetic oligoribonucleotides used to probe the interaction of MS2 coat protein with the translational operator of the MS2 replicase gene shows that the complex with the 5-BrU operator is more stable than the -5C variant; a result which is consistent with the formation of a Michael adduct at the - 5 position.
Abstract: Synthetic oligoribonucleotides have been used to probe the interaction of MS2 coat protein with the translational operator of the MS2 replicase gene. We have investigated the possible formation of a transient covalent bond between the single-stranded uridine residue, at position -5, and a cysteine side-chain on the coat protein, by the incorporation of a chemically modified residue (5-BrU) at this position. This chemically synthesised operator variant has a binding constant of between 10 and 50 times greater than that of the wild type and is therefore comparable with the tight binding variant having a cytidine substituted at the -5 position. Dissociation kinetics show that the complex with the 5-BrU operator is more stable than the -5C variant; a result which is consistent with the formation of a Michael adduct at the -5 position. In addition, a number of other chemical variants of the operator have been analysed. These include operators incorporating deoxyadenine residues at each of the important single-stranded adenine sites. Recently the Michael adduct proposal has been challenged on the basis of mutagenesis of the coat protein cysteine residues. These results are discussed in the light of our data in support of Michael adduct formation.
65 citations
••
TL;DR: The 12 half-cystines of NS1 proteins are absolutely conserved among flaviviruses, suggesting their importance to the structure and function of these proteins, and provides the basis for further refinement of flavivirus NS1 protein structural models.
65 citations
Authors
Showing all 1521 results
Name | H-index | Papers | Citations |
---|---|---|---|
Richard A. Gibbs | 172 | 889 | 249708 |
Friedrich C. Luft | 113 | 1095 | 47619 |
Alexander N. Glazer | 71 | 208 | 21068 |
Vineet Bafna | 68 | 236 | 42574 |
Kevin R. Coombes | 63 | 308 | 23592 |
Darryl J. Pappin | 61 | 170 | 29409 |
Mark D. Johnson | 60 | 289 | 16103 |
György Marko-Varga | 56 | 409 | 12600 |
Paul Thomas | 56 | 128 | 44810 |
Gerald Zon | 55 | 256 | 11126 |
Michael W. Hunkapiller | 51 | 130 | 29756 |
Bjarni V. Halldorsson | 51 | 145 | 13180 |
David H. Hawke | 50 | 157 | 9824 |
Ellson Y. Chen | 50 | 71 | 28836 |
Sridhar Hannenhalli | 49 | 162 | 21959 |