2025/09/15 by Jyoti P. Gurung, Pietro Ridone, Anaïs Biquet‐Bisquert +4 · 1 voice
Biochemistry, Genetics and Molecular Biology · Neuroscience · #Bacterial Genetics and Biotechnology #Lipid Membrane Structure and Behavior #Photoreceptor and optogenetics research
paper · pdf · doi:10.1002/mbo3.70056
openalex publication_date 2025/09/15 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/29
protein complex. Two periplasmic domains of MotB are critical in activating BFM rotation: (1) the peptidoglycan (PG) binding domain that anchors MotB in the PG layer and (2) the plug domain that modulates the proton flow. Existing cytoplasmic fluorescent probes have been shown to negatively affect motor rotation and switching. Here, we inserted a fluorescent probe in the periplasm near the plug of MotB to circumvent issues with cytoplasmic probes and for possible use in observing the mechanism of plug-based regulation of proton flow. We inserted green fluorescent protein and improved light-oxygen-voltage (LOV), a fluorescent version of the LOV domain, in four periplasmic locations in MotB. Insertions near the plug retained motility but showed limited fluorescence for both fluorophores. Additional short, flexible glycine-serine linkers improved motility but did not improve brightness. Further optimization is necessary to improve the fluorescence of these periplasmic probes.