Selective neurotensin-derived internally quenched fluorogenic substrates for neurolysin (EC 3.4.24.16): comparison with thimet oligopeptidase (EC 3.4.24.15) and neprilysin (EC 3.4.24.11).
AuthorsOliveira V, Campos M, Hemerly JP, Ferro ES, Camargo AC, Juliano MA, Juliano L
JournalAnal Biochem
PubMed ID11355859
'Internally quenched fluorescent peptides derived from neurotensin (pELYENKPRRPYIL) sequence were synthesized and assayed as substrates for neurolysin (EC 3.4.24.16), thimet oligopeptidase (EC 3.4.24.15 or TOP), and neprilysin (EC 3.4.24.11 or NEP). Abz-LYENKPRRPYILQ-EDDnp (where EDDnp is N-(2,4-dinitrophenyl)ethylenediamine and Abz is ortho-aminobenzoic acid) was derived from neurotensin by the introduction of Q-EDDnp ... More
Novel FRET-Based Assay to Detect Reverse Transcriptase Activity Using Modified dUTP Analogues.
AuthorsKrebs JF, Kore AR,
JournalBioconjug Chem
PubMed ID18163534
We have developed a novel continuous assay to measure reverse transcriptase (RT) polymerase activity. The assay uses fluorescence energy transfer measurements to detect the incorporation of complementary pairs of fluorescently labeled deoxyuridine into cDNA product. The fluorescently labeled dUTP substrates were prepared using commercially available reagents with a simple coupling ... More
Sequence verification by hybridisation with fluorescent octanucleotides as a first step to a fluorescent sequencing by hybridisation protocol.
AuthorsEickhoff H, Birch-Hirschfeld E, Scheef J, Hoyer C, Drexhage KH, Greulich KO
JournalJ Biochem Biophys Methods
PubMed ID8773548
Three sets of partly overlapping octanucleotides are 5' labelled with derivates of the fluorescence dyes fluorescein-, coumarine- and rhodamine, respectively. Hybridisation conditions are determined, under which all octanucleotides hybridise correctly against complementary target sequences bound on nylon membranes. Target sequences are three synthetic 48-mer oligonucleotides and herring sperm DNA, a ... More
Synthesis and use of an in-solution ratiometric fluorescent viscosity sensor.
AuthorsFischer D, Theodorakis EA, Haidekker MA
JournalNat Protoc
PubMed ID17401358
A procedure for the synthesis of a ratiometric viscosity fluorescent sensor is described in this protocol. The essential requirement for the design of this sensor is the attachment of a primary fluorophore that has both a viscosity-independent fluorescence emission (coumarin dye shown in blue) and an emission from a fluorophore ... More
Glutathione S-transferase P1-1 (GSTP1-1) inhibits c-Jun N-terminal kinase (JNK1) signaling through interaction with the C terminus.
AuthorsWang T, Arifoglu P, Ronai Z, Tew KD
JournalJ Biol Chem
PubMed ID11279197
c-Jun N-terminal kinase (JNK)-mediated cell signaling pathways are regulated endogenously in part by protein-protein interactions with glutathione S-transferase P1-1 (GSTP1-1) (). Using purified recombinant proteins, combined with fluorescence resonance energy transfer technology, we have found that the C terminus of JNK is critical to the interaction with GSTP1-1. The apparent ... More
Flow injection analysis of binding reaction between fluorescent lectin and cells.
AuthorsOda Y, Kinoshita M, Nakayama K, Ikeda S, Kakehi K
JournalAnal Biochem
PubMed ID10221994
A fluorometric binding assay for lectin and yeast cells using the avidin-biotin system was previously reported (Y. Oda, M. Kinoshita, and K. Kakehi, Anal. Biochem. 254, 41-48, 1997). However, the true amount of bound lectin could not be determined by this method due to difficulty in determination of the number ... More
Fluorescence studies suggest a role for alpha-synuclein in the phosphatidylinositol lipid signaling pathway.
AuthorsNarayanan V, Guo Y, Scarlata S
JournalBiochemistry
PubMed ID15641770
Alpha-synuclein plays a key role in the pathogenesis of many neurodegenerative diseases. To date, its cellular role has yet to be determined, although it has been proposed to be connected to calcium and G protein-mediated dopamine signaling. Alpha-synuclein is known to bind strongly to model membrane surfaces where it may ... More