Lanthanide-binding tags as luminescent probes for studying protein interactions.
AuthorsSculimbrene BR, Imperiali B
JournalJ Am Chem Soc
PubMed ID16734490
'Herein, we report a method for studying protein-peptide interactions which exploits the luminescence properties of Tb(III). Lanthanide-binding tags (LBTs) are short peptide sequences comprising 15-20 naturally occurring amino acids that bind Tb(III) with high affinity. These genetically encodable luminescent tags are smaller in size than the Aequorea victoria fluorescent proteins ... More
Air-stable G protein-coupled receptor microarrays and ligand binding characteristics.
AuthorsFang Y, Peng J, Ferrie AM, Burkhalter RS
JournalAnal Chem
PubMed ID16383322
'This paper described novel strategies to achieve air-stable G protein-coupled receptor (GPCR) microarrays and the uses of the microarrays for ligand profiling. Specifically, GPCR cell membrane fragments were suspended in a buffered solution containing bovine serum albumin (BSA) and disaccharide sucrose or trehalose and used for fabricating GPCR microarrays. During ... More
Multicolor in vitro translation.
AuthorsTraverso G, Diehl F, Hurst R, Shuber A, Whitney D, Johnson C, Levin B, Kinzler KW, Vogelstein B
JournalNat Biotechnol
PubMed ID12910244
'In vitro translation is a widely used tool for both analytical and preparative purposes. For analytical purposes, small amounts of proteins are synthesized and visualized by detection of labeled amino acids incorporated during translation. The original strategy of incorporating radioactively labeled amino acids, such as [35S]methionine or [14C]leucine, has been ... More
Impact of a red-shifted dye label for high throughput fluorescence polarization assays of G protein-coupled receptors.
AuthorsBanks P, Gosselin M, Prystay L
JournalJ Biomol Screen
PubMed ID11080691
'High throughput screening fluorescence polarization assays using G protein-coupled receptors (GPCRs) as targets have been compared using fluorescein and BODIPY TMR-labeled peptides. The red-shifted BODIPY TMR dye exhibits improved assay performance relative to fluorescein due to improvement in both ligand affinity to the GPCRs and assay precision brought about by ... More
Considerations for using fluorescence polarization in the screening of G protein-coupled receptors.
AuthorsBanks P, Harvey M
JournalJ Biomol Screen
PubMed ID12006109
Fluorescence polarization must be classified as a unique detection method relative to other intensity-based methods because both bound and unbound tracer is measured. Other fluorescence techniques require either physical removal of unbound tracer or a means for distinguishing only bound tracer. The presence of unbound tracer in the detection volume ... More
Solid-phase oligonucleotide synthesis and flow cytometric analysis with microspheres encoded with covalently attached fluorophores.
AuthorsNanthakumar A, Pon RT, Mazumder A, Yu S, Watson A
JournalBioconjug Chem
PubMed ID10725106
A novel combinatorial approach to synthesize oligonucleotides on fluorescently encoded microspheres based on flow sorting and segmental solid-phase synthesis is described. BODIPY dyes were covalently attached to polystyrene (8.8 microm, 55% DVB) microsphere particles to generate four fluorescently encoded sets. 20-mer oligonucleotide sequences can be synthesized on these microspheres with ... More
Fraying and electron autodetachment dynamics of trapped gas phase oligonucleotides.
AuthorsDanell AS, Parks JH
JournalJ Am Soc Mass Spectrom
PubMed ID14652182
Sensitive methods recently developed to measure laser-induced fluorescence from trapped ions have been applied to study the dynamics of double- and single-stranded oligonucleotides. In this paper, the fraying of duplex terminal base pairs has been identified by measuring the donor fluorescence as a function of temperature from an oligonucleotide duplex ... More
Fluorescence data analysis on gel-based biochips.
AuthorsBarsky V, Perov A, Tokalov S, Chudinov A, Kreindlin E, Sharonov A, Kotova E, Mirzabekov A
JournalJ Biomol Screen
PubMed ID12097187
A series of biochip readers developed for gel-based biochips includes three imaging models and a novel nonimaging biochip scanner. The imaging readers, ranging from a research-grade versatile reader to a simple portable one, use wide-field objectives and 12-bit digital large-coupled device cameras for parallel addressing of multiple array elements. This ... More
Direct, ultrasensitive, and selective optical detection of protein toxins using multivalent interactions.
AuthorsSong X, Swanson BI
JournalAnal Chem
PubMed ID10366891
Three highly sensitive, selective, and reagent-free optical signal transduction methods for detection of polyvalent proteins have been developed by directly coupling distance-dependent fluorescence self-quenching and/or resonant-energy transfer to the protein-receptor binding events. The ganglioside GM1, as the recognition unit for cholera toxin (CT), was covalently labeled with fluorophores and then ... More
Use of fluorescence polarization detection for the measurement of fluopeptidetm binding to G protein-coupled receptors.
AuthorsGagne A, Banks P, Hurt SD
JournalJ Recept Signal Transduct Res
PubMed ID12503625
G protein-coupled receptors (GPCRs) represent the single largest molecular target of therapeutic drugs currently on the market, and are also the most common target in high throughput screening assays designed to identify potential new drug candidates. A large percentage of these assays are now formatted as radioligand binding assays. Fluorescence ... More