Journal: bioRxiv

Article Title: Selective Hydrolytic Defluorination of Branched Perfluorooctanoic Acid Isomers by a Haloacid Dehalogenase

doi: 10.64898/2026.04.19.719434

Figure Lengend Snippet: (a) Docking model showing PFOA bound within the active-site pocket of 4A. (b) Enlarged view of the PFOA-binding site, illustrating predicted hydrogen-bonding interactions between the PFOA carboxylate group and surrounding active-site residues. (c) Two-dimensional interaction map of the PFOA–4A complex, highlighting hydrogen-bonding interactions involving the carboxylate group (green dashed lines) and hydrophobic or polar contacts between PFOA and active-site residues. (d) Biosensor-based plate assay detecting fluoride release following high-concentration, scaled-up reactions. PFOA (0.5 mM) was incubated with purified 4A (500 µM) at 20℃ for 120 h prior to analysis. Following incubation, aliquots from the reaction mixture were analyzed using a fluoride-responsive riboswitch biosensor with 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-gal) as the chromogenic substrate. Enzyme-only, substrate-only, buffer-only controls, and fluoride standards (10 and 500 μM KF) are shown.

Article Snippet: 5-Bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-gal) solution (20 mg/mL; Thermo Fisher Scientific, RO941) and 10 mM Tris buffer (pH 8.0, prepared from a 1 M stock; Thermo Fisher Scientific, AM9855G) were used for enzyme assays.

Techniques: Binding Assay, Concentration Assay, Incubation, Purification

Journal: The EMBO Journal

Article Title: Multifunctional roles of Brl1-Brr6 in nuclear envelope fusion during nuclear pore complex biogenesis

doi: 10.1038/s44318-026-00718-y

Figure Lengend Snippet: ( A ) AαH predictions for the wild-type and mutant sequences of Brr6 were generated using HeliQuest (Gautier et al, ). The upper cartoon illustrates the domain organization of Brr6, highlighting the two transmembrane (TM) domains, the AαH, and the four conserved cysteine residues. Amino acid positions are indicated. ( B ) Subcellular localization of GFP-tagged Brr6 wild-type and brr6 L145E AαH region. The pGal1 promoter was expressed for 3 h by the addition of galactose. Fluorescence microscopy was performed to assess localization. dsRed-HDEL was used as NE and ER marker (Madrid et al, ). Size bar: 5 µm. Representative image from three independent repeats. ( C ) Growth assay of the BRR6 shuffle strain carrying the empty LEU2 -based plasmid pRS315 or pRS315 containing the indicated BRR6 alleles ( BRR6 , brr6 L145E , and brr6 F152E ). Tenfold serial dilutions were spotted onto SC–LEU and 5-FOA plates and incubated at 30 °C for 2 days. Growth on 5-FOA plates selects for cells that have lost the URA3 plasmid, thereby testing whether the LEU2 -based plasmid alone can support viability in the absence of wild-type BRR6 . One representative of three independent experiments is shown. ( D ) Wild-type yeast cells carrying the indicated pGal1 plasmids were spotted in tenfold serial dilutions onto glucose (Glu) or galactose/raffinose (Gal/Raf) plates and incubated at 30 °C. One representative of three independent experiments is shown.

Article Snippet: Cells overexpressing these mutant alleles displayed severe growth defects on the inducing galactose plates (Gal/Raf) while control cells (pGal1- BRR6 and pGal1 vector) showed robust growth (Fig. ).

Techniques: Mutagenesis, Generated, Fluorescence, Microscopy, Marker, Growth Assay, Plasmid Preparation, Incubation