Extending the applicability of carboxyfluorescein in solid-phase synthesis.
AuthorsFischer R, Mader O, Jung G, Brock R
JournalBioconjug Chem
PubMed ID12757391
'Optimized coupling protocols are presented for the efficient and automated generation of carboxyfluorescein-labeled peptides. Side products, generated when applying earlier protocols for the in situ activation of carboxyfluorescein, were eliminated by a simple procedure, yielding highly pure fluorescent peptides and minimizing postsynthesis workup. For the cost-efficient labeling of large compound ... More
Pathway for polyarginine entry into mammalian cells.
AuthorsFuchs SM, Raines RT,
JournalBiochemistry
PubMed ID14992581
Cationic peptides known as protein transduction domains (PTDs) provide a means to deliver molecules into mammalian cells. Here, nonaarginine (R(9)), the most efficacious of known PTDs, is used to elucidate the pathway for PTD internalization. Although R(9) is found in the cytosol as well as the nucleolus when cells are ... More
Insertion and organization within membranes of the delta-endotoxin pore-forming domain, helix 4-loop-helix 5, and inhibition of its activity by a mutant helix 4 peptide.
AuthorsGerber D, Shai Y
JournalJ Biol Chem
PubMed ID10811807
The pore-forming domain of Bacillus thuringiensis Cry1Ac insecticidal protein comprises of a seven alpha-helix bundle (alpha1-alpha7). According to the "umbrella model," alpha4 and alpha5 helices form a hairpin structure thought to be inserted into the membrane upon binding. Here, we have synthesized and characterized the hairpin domain, alpha4-loop-alpha5, its alpha4 ... More
A 'puff and advance' technique for visually controlled staining of turtle retinal ganglion cells.
AuthorsAmmermüller J, Guiloff G, Normann R, Kolb H
JournalJ Neurosci Methods
PubMed ID1696675
We describe a 'puff and advance' technique for visually controlled staining of retinal ganglion cells (GCs) in the unfixed, living retina for light and electron microscopy. Glass microelectrodes are filled with rhodamine-isothiocyanate labeled horseradish peroxidase (Rh-HRP), or Lucifer yellow (LY), or a mixture of both, or with 5,6-carboxytetramethylrhodamine (5,6-Rh) and ... More
High-resolution liquid chromatography of fluorescent dye-labeled nucleic acids.
Using 100 mM of triethylammonium acetate as ion-pairing reagent, phosphodiester oligonucleotides labeled fluorescently at their 5' terminus could be separated successfully on alkylated nonporous 2.3-microns poly(styrene-divinylbenzene) particles by means of high-resolution liquid chromatography. Applying excitation wavelengths of 490, 520, 550, and 575 nm, respectively, optimum sensitivity was achieved for the ... More
Fluorescent rhodol derivatives: versatile, photostable labels and tracers.
AuthorsWhitaker JE, Haugland RP, Ryan D, Hewitt PC, Haugland RP, Prendergast FG
JournalAnal Biochem
PubMed ID1481981
A series of chemically reactive, fluorescent rhodol derivatives was prepared and evaluated. Reactive functional groups included activated esters, amines, haloacetamides, fixable hydrazide derivatives, acrylamides, and photoaffinity reagents. Depending on the choice of substituents, absorption maxima of the dyes varied from 490 to 550 nm with extinction coefficients that were generally ... More
Permeability properties of cell-to-cell channels: kinetics of fluorescent tracer diffusion through a cell junction.
AuthorsZimmerman AL, Rose B
JournalJ Membr Biol
PubMed ID4032457
We have analyzed the intracellular and cell-to-cell diffusion kinetics of fluorescent tracers in the Chironomus salivary gland. We use this analysis to investigate whether membrane potential-induced changes in junctional permeability are accompanied by changes in cell-to-cell channel selectivity. Tracers of different size and fluorescence wavelength were coinjected into a cell, ... More
A search for site-filling ligands in the Mcg Bence-Jones dimer: crystal binding studies of fluorescent compounds.
AuthorsEdmundson AB, Ely KR, Herron JN
JournalMol Immunol
PubMed ID6431268
In trigonal crystals grown in 1.9 M ammonium sulfate buffered at pH 6.2, the Mcg light-chain (Bence-Jones) dimer has a highly aromatic binding cavity accessible to a wide range of hydrophobic and aromatic ligands. A search was made for site-filling ligands by diffusing compounds into the crystals and determining their ... More