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J. Biochem, 1982, Vol. 92, No. 4 1333-1336
© 1982 Japanese Biochemical Society

other

Acridine Orange as a Fluorescent Probe for Lysosomal Proton Pump1

Yoshinori MORIYAMA, Tatsuya TAKANO and Shoji OHKUMA

Department of Microbiology and Molecular Pathology, Faculty of Pharmaceutical Sciences, Teikyo University Sagamiko, Kanagawa 199-01

Acridine orange was found to accumulate in pure lysosomal particles (tritosomes) in vitro, and the quenching of its fluorescence correlated well with the {Delta}pH (inside acid) across the lysosomal membrane. Use of this dye showed that Mg-ATP caused lysosomal acidification. This acidification was sensitive to N,N'-dicyclohexylcar-bodiimide, N-ethylmaleimide, and azide, but not to oligomycin, ouabain or vanadate. These results supported the idea of the existence of a lysosomal H+-pump, suggested in a previous paper (Ohkuma et al. (1982) Proc. Natl. Acad. Sci. U.S. 79, 2758–2762).

1This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.


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