The flexibility of actin filaments as revealed by fluorescence resonance energy transfer. The influence of divalent cations.
AuthorsNyitrai M, Hild G, Belágyi J, Somogyi B
JournalJ Biol Chem
PubMed ID10224049
'The temperature profile of the fluorescence resonance energy transfer efficiency normalized by the fluorescence quantum yield of the donor in the presence of acceptor, f'', was measured in a way allowing the independent investigation of (i) the strength of interaction between the adjacent protomers (intermonomer flexibility) and (ii) the flexibility ... More
Transglutaminase-mediated N- and C-terminal fluorescein labeling of a protein can support the native activity of the modified protein.
AuthorsTaki M, Shiota M, Taira K
JournalProtein Eng Des Sel
PubMed ID15047907
'Fluorescein and its analogs are among the best fluorophores to label proteins and the labeling generally involves chemical modification of a translated protein. Using this methodology, labeling at a specific position remains difficult. It is known that the guinea pig liver transglutaminase (TGase)-catalyzed enzymatic modification method can allow terminal-specific fluorophore ... More
Induction of tissue transglutaminase by dexamethasone: its correlation to receptor number and transglutaminase-mediated cell death in a series of malignant hamster fibrosarcomas.
AuthorsJohnson TS, Scholfield CI, Parry J, Griffin M
JournalBiochem J
PubMed ID9512467
'Treatment of the hamster fibrosarcoma cell lines (Met B, D and E) and BHK-21 hamster fibroblast cells with the glucocorticoid dexamethasone led to a powerful dose-dependent mRNA-synthesis-dependent increase in transglutaminase activity, which can be correlated with dexamethasone-responsive receptor numbers in each cell line. Increasing the number of dexamethasone-responsive receptors by ... More
Fluorescence energy transfer detects changes in fibronectin structure upon surface binding.
AuthorsWolff C, Lai CS
JournalArch Biochem Biophys
PubMed ID2913946
'We report here the changes in intramolecular distances in human plasma fibronectin (Fn) detected, upon adsorption of the protein to the surface of the Cytodex dextran microcarrier, using a fluorescence energy transfer technique. The glutamine-3 residue, near the amino terminus of each chain, was labeled enzymatically with either monodansylcadaverine (dansyl) ... More
Regulated expression of tissue transglutaminase in Swiss 3T3 fibroblasts: effects on the processing of fibronectin, cell attachment, and cell death.
AuthorsVerderio E, Nicholas B, Gross S, Griffin M
JournalExp Cell Res
PubMed ID9511731
'Tissue transglutaminase (tTgase) catalyzes the posttranslational modification of proteins by forming Ca2(+)-dependent intermolecular epsilon (gamma-glutamyl) lysine crosslinks; however, its physiological function is unclear despite increasing evidence for its involvement in the extracellular environment. To define where the enzyme is active and characterizes targets of crosslinking we have modulated tTgase expression ... More
New colored and fluorescent amine substrates for activated fibrin stabilizing factor (Factor XIIIa) and for transglutaminase.
AuthorsLorand L, Parameswaran KN, Velasco PT, Hsu LK, Siefring GE
JournalAnal Biochem
PubMed ID6193738
'Two fluorescent (FITC and 6-chloro-2-methoxyacridine) and an intensely colored (dabsyl) derivative of cadaverine were synthesized, following earlier work from this laboratory with dansyl-cadaverine, in order to enlarge the scope of possibilities for labeling some gamma-glutamine sites in proteins. Enzyme affinities of the amine substrates for human Factor XIIIa and for ... More
Evidence that the two amino termini of plasma fibronectin are in close proximity: a fluorescence energy transfer study.
AuthorsWolff C, Lai CS
JournalBiochemistry
PubMed ID3390446
'A fluorescence energy transfer technique has been used to study the intramolecular distance between the two amino termini of human plasma fibronectin. The glutamine-3 residue near the amino terminus of each chain was labeled enzymatically with either monodansylcadaverine or monofluoresceinylcadaverine by use of coagulation factor XIIIa. The nonradiative fluorescence energy ... More
The use of Fluoresceincadaverine for detecting amine acceptor protein substrates accessible to active transglutaminase in living cells.
AuthorsLajemi M, Demignot S, Borge L, Thenet-Gauci S, Adolphe M
JournalHistochem J
PubMed ID9347356
'The use of Fluoresceincadaverine as a primary amine donor for detecting the endogenous substrates for active transglutaminase in living cells was studied. Fluoresceincadaverine was found to be suitable for labelling cells in culture as it did not induce cytotoxicity when used at 0.5 mM in culture media and diffused throughout ... More
Receptor binding of fluorescein labeled steroids.
AuthorsDaxenbichler G, Grill HJ, Domanig R, Moser E, Dapunt O
JournalJ Steroid Biochem
PubMed ID7392626
A fluorescent derivative of methotrexate as an intracellular marker for dihydrofolate reductase in L1210 cells.
AuthorsHenderson GB, Russell A, Whiteley JM
JournalArch Biochem Biophys
PubMed ID6772112
The effect of phalloidin and jasplakinolide on the flexibility and thermal stability of actin filaments.
AuthorsVisegrády B, Lorinczy D, Hild G, Somogyi B, Nyitrai M
JournalFEBS Lett
PubMed ID15135072
In this work the effect of phalloidin and jasplakinolide on the dynamic properties and thermal stability of actin filaments was studied. Temperature dependent fluorescence resonance energy transfer measurements showed that filaments of Ca-actin became more rigid in the presence of phalloidin or jasplakinolide. Differential scanning calorimetric data implied that the ... More
Microchip electrophoretic immunoassay for serum cortisol.
AuthorsKoutny LB, Schmalzing D, Taylor TA, Fuchs M
JournalAnal Chem
PubMed ID8779431
An immunoassay performed using a microchip electrophoretic system is described. Separation and quantitation of free and bound labeled antigen in a competitive assay are carried out in channels micromachined into fused silica substrates. Such microchips are attractive because of their small size, ruggedness, and amenability to automated handling. The assay ... More
Capillary electrophoresis-based immunoassay for cortisol in serum.
AuthorsSchmalzing D, Nashabeh W, Yao XW, Mhatre R, Regnier FE, Afeyan NB, Fuchs M
JournalAnal Chem
PubMed ID7893004
A competitive immunoassay for cortisol based on capillary electrophoresis (CE) and laser-induced fluorescence is described. The work involved the production of assay reagents and the development of separation conditions allowing for routine analysis of serum samples. Fluorescein-labeled cortisol was synthesized and purified. Fab fragments were produced from mouse monoclonal anticortisol ... More
Fluorescence energy transfer as a probe for nucleic acid structures and sequences.
AuthorsMergny JL, Boutorine AS, Garestier T, Belloc F, Rougée M, Bulychev NV, Koshkin AA, Bourson J, Lebedev AV, Valeur B
JournalNucleic Acids Res
PubMed ID8152922
The primary or secondary structure of single-stranded nucleic acids has been investigated with fluorescent oligonucleotides, i.e., oligonucleotides covalently linked to a fluorescent dye. Five different chromophores were used: 2-methoxy-6-chloro-9-amino-acridine, coumarin 500, fluorescein, rhodamine and ethidium. The chemical synthesis of derivatized oligonucleotides is described. Hybridization of two fluorescent oligonucleotides to adjacent ... More
Using bifunctional polymers presenting vancomycin and fluorescein groups to direct anti-fluorescein antibodies to self-assembled monolayers presenting d-alanine-d-alanine groups.
AuthorsMetallo SJ, Kane RS, Holmlin RE, Whitesides GM
JournalJ Am Chem Soc
PubMed ID12683824
This paper describes the synthesis of bifunctional polyacrylamides containing pendant vancomycin (Van) and fluorescein groups, and the use of these polymers to direct antibodies against fluorescein to self-assembled monolayers (SAMs) presenting d-alanine-d-alanine (dAdA) groups. These polymers bind biospecifically to these SAMs via interactions between the dAdA and Van groups and ... More
Synthesis of a fluorescent derivative of amethopterin,.
AuthorsGapski GR, Whiteley JM, Rader JI, Cramer PL, Henderson GB, Neef V, Huennekens FM
JournalJ Med Chem
PubMed ID807734
Fluorescein isothiocyanate was treated with excess diaminopentane and the remaining unsubstituted amino group of the product was condensed, via a carbodiimide-promoted reaction, with a carboxyl group of amethopterin. The final product, a fluorescent derivative of amethopterin, was isolated by chromatography on AE-cellulose and preparative electrophoresis on polyacrylamide. It was shown ... More
Major histocompatibility complex independent T cell receptor-antigen interaction: functional analysis using fluorescein derivatives.
We have isolated T cell receptor (TCR) cDNAs from fluorescein (FL)-specific human T cell clones (alpha FL beta FL), and transferred them to TCR beta- Jurkat cells in order to study direct FL-binding to the TCR. Using either FL-conjugated polymers (FL-polymer) or FL-substituted Sepharose beads, we are able to demonstrate ... More
Replacement of ATP with ADP affects the dynamic and conformational properties of actin monomer.
AuthorsGaszner B, Nyitrai M, Hartvig N, Köszegi T, Somogyi B, Belágyi J
JournalBiochemistry
PubMed ID10504259
The effect of the replacement of ATP with ADP on the conformational and dynamic properties of the actin monomer was investigated, by means of electron paramagnetic resonance (EPR) and fluorescence spectroscopic methods. The measurement of the ATP concentration during these experiments provided the opportunity to estimate the time dependence of ... More
Interaction of recombinant procollagen and properdin modules of thrombospondin-1 with heparin and fibrinogen/fibrin.
AuthorsPanetti TS, Kudryk BJ, Mosher DF
JournalJ Biol Chem
PubMed ID9867861
Many properties have been assigned to the procollagen and properdin (Type I) modules of thrombospondin-1 (TSP1) based on activities of large proteolytic fragments of TSP1 or peptides containing TSP1-derived sequences. To examine the activities of the modules more exactly, we expressed the first properdin module (P1); the third properdin module ... More
Detection of in situ activation of transglutaminase during apoptosis: correlation with the cell cycle phase by multiparameter flow and laser scanning cytometry.
AuthorsGrabarek J, Ardelt B, Kunicki J, Darzynkiewicz Z
JournalCytometry
PubMed ID12357464
BACKGROUND: One of the hallmarks of apoptosis is activation of tissue transglutaminase (Tgase; also called transglutaminase type 2 [TGase 2]). Its activation causes cross-linking of cytoplasmic proteins, making them insoluble and presumably less immunogenic. Several biochemical and cytochemical methods to detect activity of TGase 2 exist, but none has been ... More
Functionalization of covalent DNA-streptavidin conjugates by means of biotinylated modulator components.
AuthorsNiemeyer CM, Ceyhan B, Blohm D
JournalBioconjug Chem
PubMed ID10502335
Covalent DNA-streptavidin conjugates are versatile biomolecular coupling reagents, since they have binding capacity for both a complementary nucleic acid and four molecules of biotin. The DNA-streptavidin hybrid molecules have been investigated for their capabilities to bind two different types of biotinylated components. Thus, (i) a functional biomolecule, e.g., a single-stranded ... More