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The Cl-/H+ antiporter ClC-7 is the primary chloride permeation pathway in lysosomes


Lysosomes are the stomachs of the cell—terminal organelles on the endocytic pathway where internalized macromolecules are degraded. Containing a wide range of hydrolytic enzymes, lysosomes depend on maintaining acidic luminal pH values for efficient function. Although acidification is mediated by a V-type proton ATPase, a parallel anion pathway is essential to allow bulk proton transport1,2. The molecular identity of this anion transporter remains unknown. Recent results of knockout experiments raise the possibility that ClC-7, a member of the CLC family of anion channels and transporters, is a contributor to this pathway in an osteoclast lysosome-like compartment, with loss of ClC-7 function causing osteopetrosis3. Several mammalian members of the CLC family have been characterized in detail; some (including ClC-0, ClC-1 and ClC-2) function as Cl--conducting ion channels4, whereas others act as Cl-/H+antiporters (ClC-4 and ClC-5)5,6. However, previous attempts at heterologous expression of ClC-7 have failed to yield evidence of functional protein, so it is unclear whether ClC-7 has an important function in lysosomal biology, and also whether this protein functions as a Cl- channel, a Cl-/H+ antiporter, or as something else entirely. Here we directly demonstrate an anion transport pathway in lysosomes that has the defining characteristics of a CLC Cl-/H+ antiporter and show that this transporter is the predominant route for Cl- through the lysosomal membrane. Furthermore, knockdown of ClC-7 expression by short interfering RNA can essentially ablate this lysosomal Cl-/H+ antiport activity and can strongly diminish the ability of lysosomes to acidify in vivo, demonstrating that ClC-7 is a Cl-/H+ antiporter, that it constitutes the major Cl- permeability of lysosomes, and that it is important in lysosomal acidification.

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Figure 1: 36 Cl - flux in native rat liver lysosomes.
Figure 2: Fluorescence monitoring of lysosomal H + and Cl - flux.
Figure 3: ClC-7 mediates lysosomal Cl - /H + antiport.
Figure 4: ClC-7 is essential for lysosomal acidification in vivo.


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We thank K. Swartz, M. Maduke, J. Diamond and R. Youle for critical readings of the manuscript; R. Brady, G. Murray and R. Puertollano-Moro for advice on lysosomes; and the members of the Mindell laboratory for discussions. This work was supported by the NINDS intramural program.

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Correspondence to Joseph A. Mindell.

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Graves, A., Curran, P., Smith, C. et al. The Cl-/H+ antiporter ClC-7 is the primary chloride permeation pathway in lysosomes. Nature 453, 788–792 (2008).

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