https://hdl.handle.net/10481/14652 Fri, 17 Apr 2026 04:33:14 GMT 2026-04-17T04:33:14Z https://hdl.handle.net/10481/108229 Anti-Kasha Fluorescence for Polarity Detection in Cellular Compartments Crovetto González, Luis; Fernández-Liencres, M. Paz; Villegas-Garcia, Elena; Henningway, Jodie; Girón González, María Dolores; Ríos, Ramón; Navarro, Amparo; Paredes Martínez, José Manuel https://hdl.handle.net/10481/108229 https://hdl.handle.net/10481/100098 Helically Chiral Hybrid Cyclodextrin Metal-Organic Framework with Circularly Polarized Luminescence Kazem-Rostami, Masoud; Orte Gutiérrez, Ángel; Ortuño Guzmán, Ana María; David, Arthur H. G.; Roy, Indranil; Miguel, Delia; Garci, Amine; Cruz, Carlos M.; Stern, Charlotte L.; Cuerva, Juan M.; Stoddart, J. Fraser Three achiral polycyclic aromatic fluorophores─namely, 1-pyrenecarboxylic acid, 9-anthracenecarboxylic acid, and perylene-3,9-dicarboxylic acid─were chosen based on their desired properties before being incorporated into the construction of a K+-carrying gamma-cyclodextrin (γ-CD)-based metal–organic framework (CD-MOF-1) and γ-CD-containing hybrid frameworks (CD-HFs). Among these fluorophores, only the pyrene-carrying one shows significant noncovalent bonding interactions with γ-CD in solution. This fluorophore is encapsulated in a CD-HF with a trigonal superstructure instead of the common cubic CD-MOF-1 found in the case of the other two fluorophores. Single-crystal X-ray diffraction analysis of the trigonal CD-HF reveals a π-stacked chiral positioning of the pyrene-carrying fluorophore inside the (γ-CD)2 tunnels and held uniformly around an enantiomorphous 32 screw axis along the c direction in the solid-state structure. This helix-like structure demonstrates an additional level of chirality over and above the point-chiral stereogenic centers of γ-CD and the axial chirality associated with the self-assembled π-stacked fluorophores. These arrangements result in specifically generated photophysical and chiroptical properties, such as the controlled emergence of circularly polarized luminescence (CPL) emission. In this manner, a complete understanding of the mechanism of chirality transfer from a chiral host (CD-HF) to an encapsulated achiral fluorophore has been achieved, an attribute which is often missing in the development of materials with CPL. This document is the Accepted Manuscript version of a Published Work that appeared in final form in the Journal of the American Chemical Society, copyright © 2022 American Chemical Society, after peer review and technical editing by the publisher. To access the final edited and published work see https://pubs.acs.org/articlesonrequest/AOR-NT9ZADBV72RDBFDSZNJY?_gl=1*1yv2wic*_ga*MTI3NjkyMTU3LjE3MjYxMzY0NTQ.*_ga_XP5JV6H8Q6*MTczNzYyMjQwNC40LjAuMTczNzYyMjQwNC42MC4wLjA https://hdl.handle.net/10481/100098 https://hdl.handle.net/10481/90322 Genetically encoded red fluorescent pH ratiometric sensor: Application to measuring pH gradient abnormalities in cystic fibrosis cells Salto González, Rafael; Girón González, María Dolores; Paredes Martínez, José Manuel Genetically encoded probes to measure in vivo pH are challenging. They must be chloride insensitive and require normalization of the transfection efficiency. Furthermore, probes that emit in the red or far red are advisable to promote in vivo use. The mBeRFP D162T fluorescent protein presents two emission bands with different pH sensitivities. When the probe was cytosolic-expressed in HeLa as control cells and CFBE41o, an epithelial cell line that carries an F508del mutation in the CFTR transporter, the ratiometric measurement between both emission bands allows us to determine the pH, demonstrating that mBeRFP D162T can be used to accurately measure the cytosolic pH differences of these cell lines. Furthermore, we have located the sensor inside or outside the lysosomal membrane to investigate the lysosomal pH gradient. In HeLa cells, our probe detected pH gradient changes under conditions known to alter lysosomal pH. In the CFBE41o cells, which mimic cystic fibrosis disease, we observed a complete loss of lysosomal acidification. Using lumacaftor, a drug that restores functioning CFTR protein, partially brings back the pH gradient. In conclusion, mBeRFP D162T is a valuable tool for measuring in vivo intracellular pH values and lysosomal pH gradient dynamics in physiological or pathological conditions. https://hdl.handle.net/10481/90322 https://hdl.handle.net/10481/88731 Photosensitizing properties and subcellular localisation of 3,4‑dihydro‑β‑carbolines harmaline and harmalol Denofrio, M. Paula; Paredes Martínez, José Manuel; Yañuk, Juan G.; Girón González, María Dolores; Salto González, Rafael; Talavera Rodríguez, Eva María; Crovetto González, Luis; Cabrerizo, Franco M. Harmaline (1) and harmalol (2) represent two 3,4-dihydro-β-carboline (DHβCs) most frequently reported in a vast number of living systems. Fundamental aspects including the photosensitizing properties, cellular uptake, as well as the cyto- and phototoxicity of 1 and 2 were investigated herein. The molecular basis underlying the investigated processes are elucidated. Data reveal that both alkaloids show a distinctive pattern of extracellular DNA photodamage. Compound 1 induces a DNA photodamage profile dominated by oxidised purines and sites of base loss (AP sites), whereas 2 mostly induces single-strand breaks (SSBs) in addition to a small extent of purine oxidative damage. In both cases, DNA oxidative damage would occur through type I mechanism. In addition, a concerted hydrolytic attack is suggested as an extra mechanism accounting for the SSBs formation photoinduced by 2. Subcellular internalisation, cyto- and phototoxicity of 1 and 2 and the corresponding full-aromatic derivatives harmine (3) and harmol (4) also showed quite distinctive patterns in a structure-dependent manner. These results are discussed in the framework of the potential biological, biomedical and/or pharmacological roles reported for these alkaloids. https://hdl.handle.net/10481/88731 https://hdl.handle.net/10481/86723 Efficient acetate sensor in biological media based on a selective Excited State Proton Transfer (ESPT) reaction Puente Muñoz, Virginia; Paredes Martínez, José Manuel; Resa Acosta, Sandra; Ortuño Guzmán, Ana María; Talavera Rodríguez, Eva María; Miguel Álvarez, Delia; Cuerva Carvajal, Juan Manuel; Crovetto González, Luis We have synthesized a new fluoride−containing xanthenic dye able to dynamically and quantitatively detect acetate anion, a biologically relevant analyte, in water. We studied deeply the photophysical properties of the compound and verified its use as an acetate probe in synthetic serum. https://hdl.handle.net/10481/86723