Is Single-Molecule Fluorescence Spectroscopy Ready To Join the Organic Chemistry Toolkit? A Test Case Involving Click Chemistry

Single molecule spectroscopy (SMS) has matured to a point where it can be used as a convenient tool to guide organic synthesis and drug discovery, particularly applicable to catalytic systems where questions related to homogeneous vs heterogeneous pathways are important. SMS can look at intimate mechanistic details that can inspire major improvements of the catalyst performance, its recovery, and reuse. Here, we use the click reaction between alkynes and azides as an example where improvements at the bench have been inspired and validated using single-molecule fluorescence spectroscopy

Examination of 4’,6-diamidino-2-phenylindole (DAPI) in Silica Gels through Fluorometry

Silica sol-gels synthesized through hydrolysis and condensation reactions via acid- and base-catalyzed procedures containing 4’,6-diamidino-2-phenylindole (DAPI) have been examined using fluorescence spectroscopy. DAPI is a fluorescent molecule that has traditionally been used in biosensors as a target molecule and a fluorescent stain known to bind strongly to the A-T rich regions of DNA. Sol-gels containing various concentrations of DAPI were dried conventionally to form xerogels or supercritically to form aerogels and then analyzed using fluorescence spectroscopy to determine the most optimal concentration of DAPI

Detection strategies for bioassays based on liquid chromatography, fluorescence spectroscopy and mass spectrometry

New detection strategies for bioassays based on liquid chromatography,\ud fluorescence spectroscopy and mass spectrometry were developed and are\ud presented within this thesis

From chlorophyll a towards bacteriochlorophyll a: Excited-state processes of modified pigments

By means of fluorescence spectroscopy and nonlinear absorption experiments, excited-state processes of the modified pigments [3-acetyl]-chlorophyll a, [31-OH]-bacteriochlorophyll a and [3-vinyl]-bacteriochlorophyll a were investigated and compared with those of chlorophyll a and bacteriochlorophyll a

Protein-mediated dethreading of a biotin-functionalised pseudorotaxane

In this article, we describe the synthesis of new biotin-functionalised naphthalene derivatives 3 and 4 and their complexation behaviour with avidin and neutravidin using a range of analytical techniques. We have shown using 2-(4prime or minute-hydroxyazobenzene)benzoic acid displacement and ITC experiments{,} that compounds 3 and 4 have the propensity to form reasonably high-affinity bioconjugates with avidin and neutravidin. We have also demonstrated using 1H NMR{,} UV-vis and fluorescence spectroscopy that the naphthalene moiety of 3 and 4 facilitates the formation of pseudorotaxane-like structures with 1 in water. We have then investigated the ability of avidin and neutravidin to modulate the complexation between 1 and 3 or 4. UV-vis and fluorescence spectroscopy has shown that in both cases the addition of the protein disrupts complexation between the naphthalene moieties of 3 and 4 with 1

Atomic fluorescence spectroscopy

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Using zeta-potential measurements to quantify peptide partition to lipid membranes

© The Author(s) 2011. This article is published with open access at Springerlink.com.Open Access: This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.Many cellular phenomena occur on the biomembranes. There are plenty of molecules (natural or xenobiotics) that interact directly or partially with the cell membrane. Biomolecules, such as several peptides (e.g., antimicrobial peptides) and proteins, exert their effects at the cell membrane level. This feature makes necessary investigating their interactions with lipids to clarify their mechanisms of action and side effects necessary. The determination of molecular lipid/water partition constants (Kp) is frequently used to quantify the extension of the interaction. The determination of this parameter has been achieved by using different methodologies, such as UV-Vis absorption spectrophotometry, fluorescence spectroscopy and ζ-potential measurements. In this work, we derived and tested a mathematical model to determine the Kp from ζ-potential data. The values obtained with this method were compared with those obtained by fluorescence spectroscopy, which is a regular technique used to quantify the interaction of intrinsically fluorescent peptides with selected biomembrane model systems. Two antimicrobial peptides (BP100 and pepR) were evaluated by this new method. The results obtained by this new methodology show that ζ-potential is a powerful technique to quantify peptide/lipid interactions of a wide variety of charged molecules, overcoming some of the limitations inherent to other techniques, such as the need for fluorescent labeling.This work was partially supported by project PTDC/QUI/ 69937/2006 from Fundação para a Ciência e Tecnologia-Ministério da Ciência, Tecnologia e Ensino Superior (FCT-MCTES, Portugal), and by Fundação Calouste Gulbenkian (Portugal). JMF and MMD also thank FCT-MCTES for grants IMM/BT/37-2010 and SFRH/BD/41750/2007, respectively