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Quinacrine is not a vital fluorescent probe for vesicular ATP storage.
Hasuzawa, Nao; Moriyama, Sawako; Wang, Lixiang; Nagayama, Ayako; Ashida, Kenji; Moriyama, Yoshinori; Nomura, Masatoshi.
Afiliación
  • Hasuzawa N; Division of Endocrinology and Metabolism, Department of Internal Medicine, Kurume University School of Medicine, 67 Asahi-machi, Kurume, 830-0011, Japan.
  • Moriyama S; Division of Endocrinology and Metabolism, Department of Internal Medicine, Kurume University School of Medicine, 67 Asahi-machi, Kurume, 830-0011, Japan.
  • Wang L; Department of Medical Biochemistry, Kurume University School of Medicine, Kurume, 830-0011, Japan.
  • Nagayama A; Division of Endocrinology and Metabolism, Department of Internal Medicine, Kurume University School of Medicine, 67 Asahi-machi, Kurume, 830-0011, Japan.
  • Ashida K; Division of Endocrinology and Metabolism, Department of Internal Medicine, Kurume University School of Medicine, 67 Asahi-machi, Kurume, 830-0011, Japan.
  • Moriyama Y; Division of Endocrinology and Metabolism, Department of Internal Medicine, Kurume University School of Medicine, 67 Asahi-machi, Kurume, 830-0011, Japan. moriyama_yoshinori@med.kurume-u.ac.jp.
  • Nomura M; Division of Endocrinology and Metabolism, Department of Internal Medicine, Kurume University School of Medicine, 67 Asahi-machi, Kurume, 830-0011, Japan.
Purinergic Signal ; 17(4): 725-735, 2021 12.
Article en En | MEDLINE | ID: mdl-34713379
Quinacrine, a fluorescent amphipathic amine, has been used as a vital fluorescent probe to visualize vesicular storage of ATP in the field of purinergic signaling. However, the mechanism(s) by which quinacrine represents vesicular ATP storage remains to be clarified. The present study investigated the validity of the use of quinacrine as a vial fluorescent probe for ATP-storing organelles. Vesicular nucleotide transporter (VNUT), an essential component for vesicular storage and ATP release, is present in very low density lipoprotein (VLDL)-containing secretory vesicles in hepatocytes. VNUT gene knockout (Vnut-/-) or clodronate treatment, a VNUT inhibitor, disappeared vesicular ATP release (Tatsushima et al., Biochim Biophys Acta Molecular Basis of Disease 2021, e166013). Upon incubation of mice's primary hepatocytes, quinacrine accumulates in a granular pattern into the cytoplasm, sensitive to 0.1-µM bafilomycin A1, a vacuolar ATPase (V-ATPase) inhibitor. Neither Vnut-/- nor treatment of clodronate affected quinacrine granular accumulation. In vitro, quinacrine is accumulated into liposomes upon imposing inside acidic transmembranous pH gradient (∆pH) irrespective of the presence or absence of ATP. Neither ATP binding on VNUT nor VNUT-mediated uptake of ATP was affected by quinacrine. Consistently, VNUT-mediated uptake of quinacrine was negligible or under the detection limit. From these results, it is concluded that vesicular quinacrine accumulation is not due to a consequence of its interaction with ATP but due to ∆pH-driven concentration across the membranes as an amphipathic amine. Thus, quinacrine is not a vital fluorescent probe for vesicular ATP storage.
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Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Quinacrina / Adenosina Trifosfato / Vesículas Secretoras / Hepatocitos Límite: Animals Idioma: En Revista: Purinergic Signal Año: 2021 Tipo del documento: Article País de afiliación: Japón

Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Quinacrina / Adenosina Trifosfato / Vesículas Secretoras / Hepatocitos Límite: Animals Idioma: En Revista: Purinergic Signal Año: 2021 Tipo del documento: Article País de afiliación: Japón