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Contemporary spectrofluorimeters comprise exciting light sources, excitation and emission monochromators, and detectors that without correction yield data not conforming to an ideal spectral response. The correction of the spectral properties of the exciting and emissi ...

D166V point mutation in the ventricular myosin regulatory light chain (RLC) is one of the causes of familial hypertrophic cardiomyopathy (FHC). We show here that the rates of cross-bridge attachment and dissociation are significantly different in isometrically contracting cardi ...

Atomic force microscopy in its single-molecule force spectroscopy mode is a nanomanipulation technique that is extensively used for the study of the mechanical properties of proteins. It is particularly suited to examine their response to stretching (i.e., molecular elasticity and ...

To execute their diverse range of biological functions, RNA molecules must fold into specific tertiary structures and/or associate with one or more proteins to form ribonucleoprotein (RNP) complexes. Single-molecule fluorescence spectroscopy is a powerful tool for the study of RNA ...

LHCII is the largest light-harvesting pigment-protein complex of plants, comprising more than half of photosynthetically active chlorophyll pigments in biosphere. Understanding relationship between the molecular structure of the complex and photophysical processes t ...

Bead assays are an emerging microbial detection technology with the capability for rapid detection of extremely low levels of viable pathogens. Such technologies are of high value in clinical settings and in the food industry. Here, we perform a bead assay for extracted 16S rRNA from Escheric ...

With advances in molecular biology and gene cloning techniques, it is now possible to selectively stimulate living cells of interest by using an external light source. This is done by transfecting the cells of interest with a plasmid carrying the channelrhodopsin (ChR2) gene. By stimulating ...

In Angiosperms, the reduction of protochlorophyllide (Pchlide) to chlorophyllide (Chlide), a penultimate reaction of chlorophyll biosynthesis, is catalyzed by a photoenzyme Pchlide oxidoreductase (POR) and completely inhibited in darkness. This reaction plays also a regu ...

The field of optical sensors has been a growing research area over the last three decades. A wide range of books and review articles has been published by experts in the field who have highlighted the advantages of optical sensing over other transduction methods. Fluorescence is by far the method most ...

The intracellular availability of all biologically required transition metal ions in bacteria, e.g., Zn, Cu, Fe, as well as the detoxification of nonbiological heavy metal pollutants, is controlled at the molecular level by a panel of metalloregulatory or “metal sensor” proteins. Metal se ...

The structural aspects of large macromolecular systems in solution can be conveniently addressed using the fluorescence resonance energy transfer (FRET) approach. FRET efficiency is the major parameter examined in such studies. However, its quantitative determination in ass ...

The kinetic mechanisms of biological reactions are predominantly addressed by spectroscopic stopped-flow or temperature-jump methods. Both the stopped-flow and the temperature-jump methods are relaxation kinetic techniques, i.e., they rely on examining the effect of pertu ...

Translocation of nucleic acid motor proteins (translocases) along linear nucleic acids can be studied by monitoring either the time course of the arrival of the motor protein at one end of the nucleic acid or the kinetics of ATP hydrolysis by the motor protein during translocation using pre-ste ...

A single technique is insufficient to characterize the properties of an RNA molecule so this Guide provides advice and suggestions for use of several spectroscopic methods applied to RNA. It begins with a discussion of design features to synthesize a suitable molecule for study, assuming th ...

Amphotericin B (AmB) is a polyene antibiotic used to treat deep-seated mycoses. Both the pharmaceutical and toxic activities of AmB depend on the molecular organization of the drug. The fluorescence of AmB has proven to be a powerful technique of studying the drug’s association state. In partic ...

Monte-Carlo simulation method is described and applied as an efficient tool to analyze experimental data in the presence of energy transfer in selected systems, where the use of analytical approaches is limited or even impossible. Several numerical and physical problems accompanyi ...

Cryogenic radiolytic reduction is one of the most straightforward and convenient methods of generation and stabilization of reactive iron–oxygen intermediates for mechanistic studies in chemistry and biochemistry. The method is based on one-electron reduction of the precur ...

Fluorescence lifetime imaging (FLI) has been used widely for measuring biomedical samples. Practical guidelines on taking successful FLI data are provided to avoid common errors that arise during the measurement. Several methods for analyzing and interpreting FLI results are also ...

Electro-optical effects are induced by external electric field pulses applied to solutions or suspensions and are recorded by various optical techniques. These effects are very useful for the characterization of macromolecular structures and their dynamics in solution. One of the ...

Gold–thiol chemistry is one of the most successful chemistries for conjugating biomolecules to surfaces, but such chemistry has not been exploited in optical-trapping experiments because of laser-induced ablation of gold. In this work, we describe a method to combine these two separate ...