title: Lysosome-associated miniSOG as a photosensitizer for mammalian cells.
abstract: Genetically encoded photosensitizers represent a promising optogenetic tool for the induction of light-controlled oxidative stress strictly localized to a selected intracellular compartment. Here we tested the phototoxic effects of the flavin-containing phototoxic protein miniSOG targeted to the cytoplasmic surfaces of late endosomes and lysosomes by fusion with Rab7. In HeLa Kyoto cells stably expressing miniSOG-Rab7, we demonstrated a high level of cell death upon blue-light illumination. Pepstatin A completely abolished phototoxicity of miniSOG-Rab7, showing a key role for cathepsin D in this model. Using a far-red fluorescence sensor for caspase-3, we observed caspase-3 activation during miniSOG-Rab7-mediated cell death. We conclude that upon illumination, miniSOG-Rab7 induces lysosomal membrane permeabilization (LMP) and leakage of cathepsins into the cytosol, resulting in caspase-dependent apoptosis.
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.|Nizhny Novgorod State Medical Academy, Nizhny Novgorod, Russia.
authors.forename: Alina P
authors.initials: AP
authors.lastname: Ryumina
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.|Nizhny Novgorod State Medical Academy, Nizhny Novgorod, Russia.
authors.forename: Ekaterina O
authors.initials: EO
authors.lastname: Serebrovskaya
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.
authors.forename: Dmitry B
authors.initials: DB
authors.lastname: Staroverov
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.
authors.forename: Olga A
authors.initials: OA
authors.lastname: Zlobovskaya
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.
authors.forename: Alexander S
authors.initials: AS
authors.lastname: Shcheglov
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.|Pirogov Russian National Research Medical University, Moscow, Russia.
authors.forename: Sergey A
authors.initials: SA
authors.lastname: Lukyanov
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.|Nizhny Novgorod State Medical Academy, Nizhny Novgorod, Russia.
authors.forename: Konstantin A
authors.initials: KA
authors.lastname: Lukyanov
mesh_terms: D053148:Caspase 3 / Q000032:analysis / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects / Q000528:radiation effects; D006367:HeLa Cells; D006801:Humans; D008027:Light; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008247:Lysosomes / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D062308:Optogenetics / Q000379:methods*; D017319:Photosensitizing Agents / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D026082:Singlet Oxygen / Q000378:metabolism / Q000494:pharmacology*; D020691:rab GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000090802:rab7 GTP-Binding Proteins
chemical_list: D008164:Luminescent Proteins; D017319:Photosensitizing Agents; D011993:Recombinant Fusion Proteins; D000090802:rab7 GTP-Binding Proteins; C000716463:rab7 GTP-binding proteins, human; D026082:Singlet Oxygen; D053148:Caspase 3; D020691:rab GTP-Binding Proteins
keywords: cell death; flavin-binding fluorescent protein; fluorescence microscopy; lysosome; miniSOG; phototoxicity