Regenerable biosensor platform: a total internal reflection fluorescence cell with electrochemical control.
AuthorsAsanov AN, Wilson WW, Oldham PB
JournalAnal Chem
PubMed ID9530005
'A new biosensor platform that provides simultaneous fluorescence detection and electrochemical control of biospecific binding has been developed and investigated using antibody-antigen and streptavidin-biotin interactions. Specifically, biotin was covalently bound to a transparent indium-tin oxide (ITO) working electrode, which also served as an integral part of a total internal reflection ... More
Effects of fluorescent probe structure on the dynamics at cysteine-34 within bovine serum albumin: evidence for probe-dependent modulation of the cybotactic region.
'We have prepared a series of bovine serum albumins (BSA) that have been site-selectively labeled at cysteine-34 with one of four different sulfhydryl-selective boron dipyrromethene difluoride (BODIPY) fluorescent probes (BODIPY FL IA, BODIPY FL C(1) IA, BODIPY 530/550 IA, and BODIPY 493/503 MB). We determine how the choice of extrinsic ... More
Peroxisomes in human fibroblasts have a basic pH.
AuthorsDansen TB, Wirtz KW, Wanders RJ, Pap EH
JournalNat Cell Biol
PubMed ID10620807
Caspase-3 sensitive signaling in vivo in apoptotic HeLa cells by chemically engineered intramolecular fluorescence resonance energy transfer mutants of green fluorescent protein.
AuthorsSuzuki M, Ito Y, Sakata I, Sakai T, Husimi Y, Douglas KT,
JournalBiochem Biophys Res Commun
PubMed ID15796904
Green fluorescent protein (UV5) was re-engineered to remove native cysteine residues, and a new cysteine was introduced near the C-terminus, approximately 20 A from the native fluorophore, for site-specific attachment of chemical fluorophores. The resultant efficient intramolecular FRET quenched GFP emission and gave a new emission band from the conjugated ... More
Subcellular compartmentalization of activation and desensitization of responses mediated by NK2 neurokinin receptors.
AuthorsVollmer JY, Alix P, Chollet A, Takeda K, Galzi JL
JournalJ Biol Chem
PubMed ID10608858
A functional fluorescent neurokinin NK2 receptor was constructed by joining enhanced green fluorescent protein to the amino-terminal end of the rat NK2 receptor and was expressed in human embryonic kidney cells. On cell suspensions, the binding of fluorescent Bodipy-labeled neurokinin A results in a saturatable and reversible decrease of NK2 ... More
A genetically engineered, protein-based optical biosensor of myosin II regulatory light chain phosphorylation.
AuthorsPost PL, Trybus KM, Taylor DL
JournalJ Biol Chem
PubMed ID8175704
Myosin II is an important motor in the contraction of smooth and striated muscle as well as in a variety of non-muscle cell motile events including cytokinesis, cortical contractions during migration of fibroblasts, and capping of receptors. Phosphorylation of the 20-kDa light chain by myosin light chain kinase is part ... More
Role of endocytosis in the internalization of spermidine-C(2)-BODIPY, a highly fluorescent probe of polyamine transport.
AuthorsSoulet D, Covassin L, Kaouass M, Charest-Gaudreault R, Audette M, Poulin R
JournalBiochem J
PubMed ID12097141
The mechanism of transmembrane polyamine internalization in mammalian cells remains unknown. A novel fluorescent spermidine conjugate [Spd-C(2)-BODIPY; N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionyl)-N'-(S -[spermidine-(N(4)-ethyl)]thioacetyl)ethylenediamine] was synthesized from N(4)-(mercaptoethyl)spermidine by a simple, one-step coupling procedure. In Chinese-hamster ovary (CHO) cells, Spd-C(2)-BODIPY accumulation was inhibited by exogenous putrescine, spermidine and spermine, was subject to feedback transport inhibition ... More
Site-directed targeting of plasminogen activator inhibitor-1 as an example for a novel approach in rational drug design.
AuthorsDe Taeye B, Compernolle G, Declerck PJ
JournalJ Biol Chem
PubMed ID14988411
As plasminogen activator inhibitor-1 (PAI-1), the physiological inhibitor of tissue-type plasminogen activator, is considered to be an important risk factor in several (patho)physiological conditions, many research activities focus on attempts to inhibit this serpin. The approach illustrated in the current study focuses on elucidating important interaction sites allowing the inhibition ... More