Reaction of cystathionase with the fluorescent probe bis(dansyl)cystine.
AuthorsOh KJ, Churchich JE
JournalJ Biol Chem
PubMed ID4846744
Characterization of the uptake and toxicity of a fluorescent thiol reagent.
AuthorsOlive PL, Biaglow JE, Varnes ME, Durand RE
JournalCytometry
PubMed ID6839884
'3-(4-Maleimidylphenyl)-4-methyl-7-diethylamino coumarin (CPM), is a fluorescent thiol-binding agent. CPM is nontoxic to Chinese hamster V-79 cells (2 x 10(5) cells/ milliliter) exposed to 2.5 micrograms/milliliter for 30 minutes. However, both toxicity and cellular binding were directly dependent on the drug:cell ratio. Using flow cytometry, cellular binding of CPM correlated with ... More
Activated polyurethane modified with latent thiol groups.
AuthorsAlferiev IS, Fishbein I
JournalBiomaterials
PubMed ID12361613
'A novel type of modified polyurethane with pendant acetylthio groups (as a latent form of thiol groups) has been proposed for the use in surface modifications with various biomolecules. The polymer was prepared via a modified variant of low-temperature bromoalkylation of urethane hard segments followed by the reaction of pendant ... More
The FMN to heme electron transfer in cytochrome P450BM-3. Effect of chemical modification of cysteines engineered at the FMN-heme domain interaction site.
'The crystal structure of the complex between the heme and FMN-containing domains of Bacillus megaterium cytochrome P450BM-3 (Sevrioukova, I. F., Li, H., Zhang, H., Peterson, J. A., and Poulos, T. L. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 1863-1868) indicates that the proximal side of the heme domain ... More
Na+- and cGMP-induced Ca2+ fluxes in frog rod photoreceptors.
AuthorsSchnetkamp PP, Bownds MD
JournalJ Gen Physiol
PubMed ID3031199
'We have examined the Ca2+ content and pathways of Ca2+ transport in frog rod outer segments using the Ca2+-indicating dye arsenazo III. The experiments employed suspensions of outer segments of truncated, but physiologically functional, frog rods (OS-IS), intact isolated outer segments (intact OS), and leaky outer segments (leaky OS with ... More
Lectin receptors of rods and cones. Visualization by fluorescent label.
AuthorsBridges CD
JournalInvest Ophthalmol Vis Sci
PubMed ID6969714
'The binding of eight fluorescence-labeled lectins to visual cells was investigated. Concanavalin A (Con A) (specific for oligosaccharides containing mannose) bound evenly to the surfaces of rod outer segments (ROSs) from frog, cattle, goldfish, and turtle (Pseudemys). In frog, binding was observed on the inner segment, with intensifications at the ... More
Comparison of the fluorescence and conformational properties of smooth and striated tropomyosin.
AuthorsLehrer SS, Betteridge DR, Graceffa P, Wong S, Seidel JC
JournalBiochemistry
PubMed ID6722112
'In contrast to previous conformational studies with rabbit skeletal and cardiac tropomyosins, (i) when the cysteine side chains of chicken gizzard tropomyosin were reacted with 5,5''-dithiobis(2-nitrobenzoate), an interchain disulfide cross-link was not produced, (ii) when they were labeled with pyrenylmaleimide , excimer fluorescence was not observed, and (iii) when they ... More
On the evaluation of photoreceptor properties by micro-fluorimetric measurements of fluorochrome diffusion.
AuthorsHochstrate P, Rüppel H
JournalBiophys Struct Mech
PubMed ID7388121
'By use of the microfluorimetric technique it is possible to study the diffusion of the fluorochrome di-dansylcystine (DDC) within isolated frog rod outer segments (ros) which are immobilysed in agarose gel. For this purpose, by a short hypotonic shock a leak is applied to one end of the ros. By ... More
The reaction of S-mercuric-N-dansylcysteine with acetylcholinesterase and butyrylcholinesterase.
AuthorsTomlinson G, Kinsch EM
JournalBiochem Cell Biol
PubMed ID2789787
'S-mercuric-N-dansylcysteine was investigated as a potential probe of protein sulphydryl groups using bovine serum albumin, S-carboxymethyl-bovine serum albumin, lysozyme, and partially reduced lysozyme as test proteins. Criteria used to assess covalent binding through mercury-bridged mercaptide linkages include a finite reaction time (minutes to hours), abolition of the characteristic fluorescence spectrum ... More
Spectroscopic studies of stellacyanin derivatives.
AuthorsKnaff DB, Harsh CE, Holwerda RA
JournalBiochemistry
PubMed ID6456761
'Two covalently modified derivatives of the apoprotein of the blue copper protein stellacyanin have been prepared. In one case, a dansyl group was linked to the cysteine at the copper binding site of apostellacyanin; in the other, a nitrophenol moiety has been attached to this same cysteine. Fluorescence yields and ... More
Formation of mixed disulfide adducts at cysteine-281 of the lactose repressor protein affects operator and inducer binding parameters.
AuthorsDaly TJ, Olson JS, Matthews KS
JournalBiochemistry
PubMed ID3535878
'The lactose repressor protein has been modified with three sulfhydryl-specific reagents which form mixed disulfide adducts. Methyl methanethiosulfonate (MMTS) and 5,5''-dithiobis(2-nitrobenzoic acid) (DTNB) completely reacted with all three cysteine residues, whereas only partial reaction was observed with didansylcystine. Cysteines-107 and -140 reacted stoichiometrically with MMTS and DTNB, while Cys-281 was ... More
Agglutination of isolated rod outer segments by lectins.
AuthorsBridges CD
JournalInvest Ophthalmol Vis Sci
PubMed ID6969711
'The interactions of ten lectins with rod outer segment (ROS) plasma membranes and the effects of various haptene sugars were examined by studying agglutination in suspensions of washed ROSs. It is concluded that agglutination sites on the ROS membrane involve receptors that are recognized by eight lectins with haptene sugar ... More
Two conformational states of didansylcystine-labeled rabbit cardiac tropomyosin.
AuthorsBetteridge DR, Lehrer SS
JournalJ Mol Biol
PubMed ID6864806
'The fluorescence properties of rabbit cardiac tropomyosin specifically labeled at Cys190 with didansylcystine has been correlated with its unfolding transitions. Circular dichroism studies at 222 nm showed that for both didansylcystine-labeled tropomyosin and reduced tropomyosin under physiological conditions, a small thermal pretransition near 30 degrees C was present before the ... More
The G-actin is greater than F-actin transformation as studied by the fluorescence of bound dansyl cystine.
AuthorsCheung HC, Cooke R, Smith L
JournalArch Biochem Biophys
PubMed ID5545487
Interaction of globular actin with myosin subfragments.
AuthorsCooke R, Morales MF
JournalJ Mol Biol
PubMed ID4329225
Conformational aspects of rhodopsin and retinal disc membranes.
AuthorsSteinemann A, Wu C, Stryer L
JournalJ Supramol Struct
PubMed ID4767463
Topology of the outer segment membranes of retinal rods and cones revealed by a fluorescent probe.
AuthorsYoshikami S, Robinson WE, Hagins WA
JournalScience
PubMed ID4138020
When N,N'-didansyl cystine binds to the cell membranes of vertebrate rods and cones its fluorescence efficiency increases about 20-fold. The entire outer segments of living cones become brilliantly fluorescent. Stained live rods, as well as most freshly detached outer segments, are only weakly fluorescent, but they become brightly fluorescent within ... More
Photoaffinity labeling of antibodies for applications in homogeneous fluoroimmunoassays.
AuthorsChang IN, Lin JN, Andrade JD, Herron JN
JournalAnal Chem
PubMed ID7762830
A homogeneous noncompetitive immunoassay based on photoaffinity labeling techniques is described. Using this method, a fluorophore (reporter) can be specifically attached to an antibody in the vicinity of its antigen-combining sites. Upon antigen binding, changes in the fluorescence spectrum of the reporter molecule are often observed. Two fluorophores, pyrene and ... More
Dietary ascorbic acid and hepatic mixed function oxidase activity in the guinea pig.
AuthorsPeterson FJ, Holloway DE, Duquette PH, Rivers JM
JournalBiochem Pharmacol
PubMed ID6830622
Studies were carried out to characterize the response of hepatic mixed function oxidase (MFO) activity to chronic ascorbic acid deficiency and excessive ascorbic acid intake in the guinea pig. When guinea pigs were fed excessive ascorbic acid, there was a small increase in hepatic cytochrome P-450 which was unaccompanied by ... More
Proximity relationships in rhodopsin.
AuthorsWu CW, Stryer L
JournalProc Natl Acad Sci U S A
PubMed ID4504322
Energy transfer was used as a spectroscopic ruler to deduce proximity relationships within bovine rhodopsin in digitonin solution. Rhodopsin was specifically labeled with fluorescent chromophores at three sites. Site A was alkylated by fluorescent derivatives of iodoacetamide. Site B was labeled by fluorescent disulfides, by a disulfide-sulfhydryl interchange reaction. Sites ... More
Flow cytometry techniques for studying cellular thiols.
AuthorsDurand RE, Olive PL
JournalRadiat Res
PubMed ID6193555
Cellular thiols, and especially glutathione, act as scavenger nucleophiles and can protect against toxicity, mutagenicity, or transformation by ionizing radiation and many carcinogens. Development of a rapid assay to quantitate the cellular content of thiols could thus be useful in assessing or predicting cellular risk to damage. Several fluorescent thiol-reactive ... More
Fluorescent localization of membrane sites in glycerinated chicken skeletal muscle fibers and the relationship of these sites to the protein composition of the Z disc.
AuthorsLazarides E, Granger BL
JournalProc Natl Acad Sci U S A
PubMed ID358193
Didansyl derivatives of amino acids and N-phenyl-1-naphthylamine were used to localize membrane hydrophobic sites in glycerol-extracted chicken skeletal muscle fibers. Epifluorescence microscopy revealed that such sites coincide with the distribution of mitochondria, the transverse tubular (T) system and the sarcoplasmic reticulum (SR). They are specifically associated with myofibril Z lines ... More