Quantification of carboxyl groups in carbodiimide cross-linked collagen sponges.
AuthorsEveraerts F, Torrianni M, Hendriks M, Feijen J,
JournalJ Biomed Mater Res A
PubMed ID17595021
'Glutaraldehyde (GA) fixation of bioprosthetic tissue is a well adapted technique, with commercial products on the market for almost 40 years. Amine groups present in tissue react with GA to form different types of cross-links. An estimation of the degree of cross-linking of the tissue can be obtained by measuring ... More
Localization of the hinge region of the Ca(2+)-ATPase of sarcoplasmic reticulum using resonance energy transfer.
AuthorsBaker KJ, East JM, Lee AG
JournalBiochim Biophys Acta
PubMed ID8204650
'The Ca(2+)-ATPase of skeletal muscle sarcoplasmic reticulum can be labelled at Cys-670 and Cys-674 with 5-[[2-[(iodoacetyl) amino]ethyl]amino]naphthalene-1-sulphonic acid (IAEDANS). Resonance energy transfer has been used to measure the distance between Cys-670/Cys-674 and Glu-439 labelled with 5-(bromomethyl)fluorescein as 40 A. The height of Cys-670/Cys-674 above the phospholipid/water interface has been measured ... More
Measurement of thiol-containing amino acids and phytochelatin (PC2) via capillary electrophoresis with laser-induced fluorescence detection.
AuthorsHart JJ, Welch RM, Norvell WA, Kochian LV,
JournalElectrophoresis
PubMed ID11824625
'An analytical method for determining thiols and phytochelatins using high-performance capillary electrophoresis coupled with laser-induced fluorescence detection is presented. The technique utilizes the labeling of thiols with the fluorescent reagent 5-bromomethylfluorescein (5-BMF), which is excited by a 488 nm argon ion laser and fluoresces at 515 nm. The paper describes ... More
Ultraviolet and fluorescence derivatization reagents for carboxylic acids suitable for high performance liquid chromatography: a review.
AuthorsMukherjee PS, Karnes HT
JournalBiomed Chromatogr
PubMed ID8879524
Pre- and postcolumn derivatization with a suitable chromophore or fluorophore have often been utilized to enhance the sensitivity and selectivity of detection for analytes possessing a carboxyl group. The major classes of UV and fluorescent derivatization reagents include the coumarin analogues, alkyl halides, diazoalkanes, and amines. The derivatization reaction conditions, ... More
Labeling the (Ca(2+)-Mg2+)-ATPase of sarcoplasmic reticulum at Glu-439 with 5-(bromomethyl)fluorescein.
AuthorsStefanova HI, Mata AM, Gore MG, East JM, Lee AG
JournalBiochemistry
PubMed ID8099497
The (Ca(2+)-Mg2+)-ATPase of skeletal muscle sarcoplasmic reticulum was labeled with 5-(bromomethyl)fluorescein. A stoichiometry of one label per ATPase molecule was found, which was unaffected by the presence of ATP. Labeling resulted in a 60% decrease in ATPase activity. Sequencing identified the labeled residue as Glu-439. The fluorescence emission spectrum of ... More
Analysis of gamma-(cholesteryloxy)butyric acid in biologic samples by derivatization with 5-(bromomethyl)fluorescein followed by high-performance liquid chromatography with laser-induced fluorescence detection.
AuthorsMukherjee PS, Karnes HT
JournalAnal Chem
PubMed ID9027240
This report describes the first application of 5-(bromo-methyl)fluorescein (5-BMF) for the quantitation of a pharmaceutically relevant carboxyl-containing analyte in a biological matrix. An analytical method for quantitation of gamma-(cholesteryloxy)butyric acid (CBA), a relatively new antitumor agent, in different tissues of Sprague-Dawley rats was developed. 5-BMF was employed to form a ... More
A fluorescence-based method for analyzing retinoic acid in biological samples.
AuthorsDonato LJ, Noy N
JournalAnal Biochem
PubMed ID16919229
Retinoic acid (RA) modulates the rates of transcription of numerous genes and thus plays key roles in multiple biological processes and is used in therapy of a number of diseases. However, RA therapy is often confounded by toxicity, raising the need for methodologies for its ready quantitation in biological samples. ... More
Evidence for long range allosteric interactions between the extracellular and cytoplasmic parts of bacteriorhodopsin from the mutant R82A and its second site revertant R82A/G231C.
AuthorsAlexiev U, Mollaaghababa R, Khorana HG, Heyn MP
JournalJ Biol Chem
PubMed ID10788455
Evidence is presented for long range interactions between the extracellular and cytoplasmic parts of the heptahelical membrane protein bacteriorhodopsin in the mutant R82A and its second site revertant R82A/G231C. (i) In the double mutants R82A/G72C and R82A/A160C, with the cysteine mutation on the extracellular or cytoplasmic surface, respectively, the photocycle ... More
Rapid long-range proton diffusion along the surface of the purple membrane and delayed proton transfer into the bulk.
AuthorsAlexiev U, Mollaaghababa R, Scherrer P, Khorana HG, Heyn MP
JournalProc Natl Acad Sci U S A
PubMed ID7831293
The pH-indicator dye fluorescein was covalently bound to the surface of the purple membrane at position 72 on the extracellular side of bacteriorhopsin and at positions 101, 105, 160, or 231 on the cytoplasmic side by reacting bromomethylfluorescein with the sulfhydryl groups of cysteines introduced by site-directed mutagenesis. At position ... More
5-Bromomethyl fluorescein (5-BMF) for derivatization of carboxyl containing analytes for use with laser-induced fluorescence detection.
AuthorsMukherjee PS, DeSilva KH, Karnes HT
JournalPharm Res
PubMed ID7667203
PURPOSE. 5-Bromomethyl fluorescein (5-BMF) was evaluated in this work as a pre-column, off-line derivatizing reagent for analytes containing a free carboxyl group. The reagent possessed high molar absorptivity and quantum yield and it's excitation maximum matched the intense 488.0 nm emission line of an argon ion laser. The thermal and ... More