The molecular chaperonin cpn60 displays local flexibility that is reduced after binding with an unfolded protein.
AuthorsGorovits BM, Horowitz PM
JournalJ Biol Chem
PubMed ID7768899
'Steady-state fluorescence polarization was used to examine the chaperonin cpn60 that was covalently labeled with pyrene. Two compounds, 1-pyrenesulfonyl chloride or N-(1-pyrene)maleimide, were used to incorporate up to 8 mol of pyrene per mol of cpn60 14-mer. The fluorescence lifetime of the cpn60-pyrenesulfonyl chloride conjugate exhibited a double exponential decay: ... More
Adenosine deaminase in cell transformation. Biophysical manifestation of membrane dynamics.
AuthorsPorat N, Gill D, Parola AH
JournalJ Biol Chem
PubMed ID2844778
'Cell transformation is associated with a dramatic collapse of a graphic fingerprint characteristic of normal cells, as measured by phase fluorimetry. This is demonstrated on adenosine deaminase (ADA, EC 3.5.4.4), an established malignancy marker. ADA activity is known to decrease markedly in chick embryo fibroblasts (CEF) transformed by Rous sarcoma ... More
Synthesis and use of labelled nucleoside phosphoramidite building blocks bearing a reporter group: biotinyl, dinitrophenyl, pyrenyl and dansyl.
AuthorsRoget A, Bazin H, Teoule R
JournalNucleic Acids Res
PubMed ID2798121
'The synthesis of protected nucleoside phosphoramidites bearing various markers such as biotinyl, dinitrophenyl, dansyl and pyrenyl groups are reported. These labelled deoxynucleosides phosphoramidites were used for solid phase oligonucleotide synthesis in the same way than the usual protected phosphoramidities without any change in the synthetic cycle and the deprotection step. ... More
Pyrenesulfonyl azide: a marker of acetylcholine receptor subunits in contact with membrane hydrophobic environment.
AuthorsSator V, Gonzalez-Ros JM, Calvo-Fernandez P, Martinez-Carrion M
JournalBiochemistry
PubMed ID427109
Interaction of a novel fluorescent analog of interferon-gamma with transformed cells.
AuthorsFalach A, Nathan I, Baram S, Porat N, Dvilansky A, Parola AH
JournalBioconjug Chem
PubMed ID9258441
A fluorescent analog of human recombinant interferon-gamma (IFN-gamma) was prepared for the first time. The recovered pyrene-labeled IFN-gamma (py-IFN-gamma), with an estimated seven pyrene molecules per IFN-gamma, retained over half of its original biological activity. Binding of py-IFN-gamma to human amnion WISH cells showed appreciable enhancement in fluorescence polarization from ... More
Enhancement of fluorescence of pyrene-containing lipids by polar media, detergents and phospholipids.
AuthorsLevade T, Salvayre R, Gatt S
JournalExperientia
PubMed ID3653339
The fluorescence intensities of a medium-chain fatty acid and of several amphiphilic lipids, each containing pyrene in covalent linkage, were enhanced considerably by: 1) Dissolving in mixtures of a polar solvent (e.g. methanol, ethanol, tetrahydrofuran or dimethylsulfoxide) and water; for each individual compound, a certain ratio of solvent to water ... More
Fluorochromes with long-lived fluorescence as potential labels for pulsed laser immunocytofluorometry: photophysical characterization of pyrene derivatives.
AuthorsAndreoni A, Bottiroli G, Colasanti A, Giangarè MC, Riccio P, Roberti G, Vaghi P
JournalJ Biochem Biophys Methods
PubMed ID7836661
An apparatus for laser-induced time-resolved fluorescence measurements, in conjunction with pyrene derivatives endowed with long-lived excited singlet states, was employed for immunocytofluorometric measurements. N-(1-pyrene)maleimide, 1-pyrenesulfonyl chloride and 1-pyreneisothiocyanate were conjugated with immunoglobulin, antimouse-IgG, and the fluorescence decays of both free and conjugated forms were investigated. Bi-exponential decays were obtained in ... More
Phospholipase A2 activities with a plasmalogen substrate in brain and in neural tumor cells: a sensitive and specific assay using pyrenesulfonyl-labeled plasmenylethanolamine.
AuthorsHirashima Y, Mills JS, Yates AJ, Horrocks LA
JournalBiochim Biophys Acta
PubMed ID2174264
We have developed a new assay method for phospholipase A2 (EC 3.1.1.4.), towards ethanolamine plasmalogen using pyrenesulfonyl-labeled plasmenylethanolamine as the substrate. This procedure is sensitive to about 3 pmol/ml per min and is absolutely specific for plasmalogen. In this method, the product of phospholipase A2, pyrenesulfonyl-labeled lysoplasmalogen, is hydrolyzed to ... More
Nanosecond fluorescence spectroscopy of human immunoglobulin A.
AuthorsLiu BM, Cheung HC, Mestecky J
JournalBiochemistry
PubMed ID7225369
The solution properties of five samples of human immunoglobulin A (IgA) were investigated with covalent and hydrophobic fluorescence probes. The immunoglobulins included a secretory IgA and four myeloma proteins of both IgA1 and IgA2 subclasses in the monomeric and dimeric forms. The probe 8-anilinonaphthalene-1-sulfonate (ANS) was found to bind to ... More
Tight binding of bulky fluorescent derivatives of adenosine to the low affinity E2ATP site leads to inhibition of Na+/K+-ATPase. Analysis of structural requirements of fluorescent ATP derivatives with a Koshland-Némethy-Filmer model of two interacting ATP sites.
AuthorsThoenges D, Amler E, Eckert T, Schoner W
JournalJ Biol Chem
PubMed ID9890953
A Koshland-Némethy-Filmer model of two cooperating ATP sites has previously been shown to explain the kinetics of inhibition of Na+/K+-ATPase (EC 3.6.1.37) by dansylated ATP (Thoenges, D., and Schoner, W. (1997) J. Biol. Chem. 272, 16315-16321). The present work demonstrates that this model adequately describes all types of interactions and ... More
Cholesterol dependent recruitment of di22:6-PC by a G protein-coupled receptor into lateral domains.
AuthorsPolozova A, Litman BJ
JournalBiophys J
PubMed ID11053136
Bovine rhodopsin was reconstituted into mixtures of didocosahexaenoylphosphatidylcholine (di22:6-PC), dipalmitoylphosphatidylcholine (di16:0-PC), sn-1-palmitoyl-sn-2-docosahexaenoylphosphatidylcholine (16:0, 22:6-PC) and cholesterol. Rhodopsin denaturation was examined by using high-sensitivity differential scanning calorimetry. The unfolding temperature was increased at lower levels of lipid unsaturation, but the highest temperature was detected for native disk membranes: di22:6-PC < 16:0,22:6-PC ... More
Pyrene-labeled DNA probes for homogeneous detection of complementary DNA sequences: poly(C) model system.
AuthorsYguerabide J, Talavera E, Alvarez JM, Afkir M
JournalAnal Biochem
PubMed ID8921193
DNA and RNA probes are important analytical reagents in molecular biology and in the detection of infectious and genetic diseases. However, the present polynucleotide probe technology is complex and labor-intensive. We have been investigating the possibility of using fluorescent-labeled DNA probes to develop assays which do not require the separation ... More
A modular strategy for tailoring fluorescent biosensors from ribonucleopeptide complexes.
AuthorsHagihara M, Fukuda M, Hasegawa T, Morii T
JournalJ Am Chem Soc
PubMed ID17002390
Fluorescent biosensors that facilitate reagentless sensitive detection of small molecules are crucial tools in the areas of therapeutics and diagnostics. However, construction of fluorescent biosensors with desired characteristics, that is, detection wavelengths and concentration ranges for ligand detection, from macromolecular receptors is not a straightforward task. An ATP-binding ribonucleopeptide (RNP) ... More
Zymogen/enzyme discrimination using peptide chloromethyl ketones.
AuthorsWilliams EB, Krishnaswamy S, Mann KG
JournalJ Biol Chem
PubMed ID2708377
Glutamylglycinylarginyl chloromethyl ketone, tyrosylglycinylarginyl chloromethyl ketone, and phenylalanylprolylarginyl chloromethyl ketone have been labeled at their amino termini using fluorescein, rhodamine-X, lissamine-rhodamine, pyrene, and the 1,5-, 2,5-, and 2,6-dimethylaminonaphthalene-1-sulfonyl moieties. These peptidyl chloromethyl ketones have also been modified by incorporation of biotin and epsilon-amino caproyl biotin. The ability of these various ... More