Förster resonance energy transfer (FRET) is an established fluorescence-based method used to successfully measure distances in and between biomolecules in vitro as well as within cells. In FRET, the efficiency of energy transfer, measured by changes in fluorescence intensity or lifetime, relates to the distance between two fluorescent molecules or labels. Determination of dynamics and conformational changes from the distances are just some examples of applications of this method to biological systems. Under certain conditions, this methodology can add to and enhance existing X-ray crystal structures by providing information regarding dynamics, flexibility, and adaptation to binding surfaces. We describe the use of FRET and associated distance determinations to elucidate structural properties, through the identification of a binding site or the orientations of dimer subunits. Through judicious choice of labeling sites, and often employment of multiple labeling strategies, we have successfully applied these mapping methods to determine global structural properties in a protein-DNA complex and the SecA-SecYEG protein translocation system. In the SecA-SecYEG system, we have used FRET mapping methods to identify the preprotein-binding site and determine the local conformation of the bound signal sequence region. This study outlines the steps for performing FRET mapping studies, including identification of appropriate labeling sites, discussion of possible labels including non-native amino acid residues, labeling procedures, how to perform measurements, and interpreting the data.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10639101PMC
http://dx.doi.org/10.3791/63433DOI Listing

Publication Analysis

Top Keywords

energy transfer
12
förster resonance
8
resonance energy
8
global structural
8
structural properties
8
labeling sites
8
mapping methods
8
fret mapping
8
fret
5
mapping
4

Similar Publications

Direct photochemical conversion of CO2 into a single carbon-based product currently represents one of the major issues in the catalysis of the CO2 reduction reaction (CO2RR). In this work, we demonstrate that the combination of an organic photosensitizer with a heptacoordinated iron(II) complex allows to attain a noble-metal-free photochemical system capable of efficient and selective conversion of CO2 into CO upon light irradiation in the presence of N,N-diisopropylethylamine (DIPEA) and 2,2,2-trifluoroethanol (TFE) as the electron and proton donor, respectively, with unprecedented performances (ΦCO up to 36%, TONCO > 1000, selectivity > 99%). As shown by transient absorption spectroscopy studies, this can be achieved thanks to the fast rates associated with the electron transfer from the photogenerated reduced dye to the catalyst, which protect the dye from parallel degradation pathways ensuring its stability along the photochemical reaction.

View Article and Find Full Text PDF

[The many ways flowers send signals to pollinators].

Biol Aujourdhui

January 2025

Institut d'Écologie et des Sciences de l'Environnement de Paris (iEES Paris), Paris, France - Sorbonne Université, 4 place Jussieu, 75005 Paris, France.

The evolutionary success of angiosperms, which make up more than 95 percent of the world's terrestrial flora, is largely based on their interactions with animal pollinators. Indeed, it is estimated that, on average, 87.5 percent of flowering plants are pollinated by animals.

View Article and Find Full Text PDF

[FeFe] hydrogenases make up a structurally diverse family of metalloenzymes that catalyze proton/dihydrogen interconversion. They can be classified into phylogenetically distinct groups denoted A-G, which differ in structure and reactivity. Prototypical Group A hydrogenases have high turnover rates and remarkable energy efficiency.

View Article and Find Full Text PDF

The primary production of fjords across the Arctic and Subarctic is undergoing significant transformations due to the climatically driven retreat of glaciers and ice sheets. However, the implications of these changes for upper trophic levels remain largely unknown. In this study, we employ both bulk and compound-specific stable isotope analyses to investigate how shifts at the base of fjord food webs impact the carbon and energy sources of consumers.

View Article and Find Full Text PDF

Along with the development of miniaturization, integration, and high power of electronic chips in the 5G and artificial intelligence era and their urgent need for technologies enabled to solve high heat flux dissipation in limited space, investigating bioinspired extreme superwettability surfaces with high-efficiency condensation heat transfer (CHT) performance has attracted great interest in academic and industrial communities. Compared with filmwise condensation of flat hydrophilic surfaces featured with continuous liquid films, dropwise condensation of flat hydrophobic surfaces is a more efficient type of energy transport way. However, discrete condensate drops can only shed off the hydrophobic flat surfaces under gravity until their sizes reach the capillary length of liquid, e.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!