AI Article Synopsis

  • - ClC-6 and ClC-7 are intracellular proteins that help regulate chloride (Cl) and hydrogen ions (H) in late endosomes and lysosomes, but they function differently, with ClC-7 needing another protein called Ostm1 to operate properly.
  • - Experiments showed that lower levels of external Cl reduce ClC-6 function, while they enhance ClC-7's activity, particularly under acidic conditions or low internal Cl levels.
  • - The study highlighted unique functions and responses of both transporters, indicating that ClC-7's ability to handle Cl may contribute to disease processes like osteopetrosis, emphasizing the need for balanced ion levels in cellular environments.

Article Abstract

ClC-6 and ClC-7 are closely related, intracellular Cl /H antiporters belonging to the CLC family of channels and transporters. They localize to acidic late endosomes and lysosomes and probably function in ionic homeostasis of these contiguous compartments. ClC-7 transport function requires association with the accessory protein Ostm1, whereas ClC-6 transport does not. To elucidate their roles in endo-lysosomes, we measured Cl - and pH-dependences of over-expressed wild-type ClC-6 and ClC-7, as well as disease-associated mutants, using high-resolution recording protocols. Lowering extracellular Cl (corresponding to luminal Cl in endo-lysosomes) reduced ClC-6 currents, whereas it increased transport activity of ClC-7/Ostm1. Low extracellular Cl activated ClC-7/Ostm 1 under acidic extracellular conditions, as well as under conditions of low intracellular chloride. Activation is conserved in ClC-7 , a variant displaying disrupted PI(3,5)P inhibition. Detailed biophysical analysis of disease-associated ClC-6 and ClC-7 gain-of-function (GoF) variants, ClC-6 and ClC-7 , and the ClC-7 and ClC-6 correlates, identified additional functional nuances distinguishing ClC-6 and ClC-7. ClC-7 recapitulated GoF produced by ClC-6 . ClC-6 displayed transport activation qualitatively similar to ClC-7 , although current density did not differ from that of wild-type ClC-6. Finally, rClC-7 , homologous to hClC-7 , an osteopetrosis variant with fast gating kinetics, appeared indifferent to extracellular Cl , identifying altered Cl sensitivity as a plausible mechanism underlying disease. Collectively, the present studies underscore the distinct roles of ClC-6 and ClC-7 within the context of their respective localization to late endosomes and lysosomes. In particular, we suggest the atypical inhibition of ClC-7 by luminal Cl serves to limit excessive intraluminal Cl accumulation. KEY POINTS: ClC-6 and ClC-7 are late endosomal and lysosomal 2 Cl /1 H exchangers, respectively. When targeted to the plasma membrane, both activate slowly at positive voltages. ClC-6 activity is decreased in low extracellular (i.e. luminal) chloride, whereas ClC-7 is activated by low luminal chloride, even at acidic pH. The functional gain-of-function phenotypes of the ClC-6 and ClC-7 disease mutations ClC-6 and ClC-7 are maintained when introduced in their respective homologues, ClC-7 and ClC-6 , with all mutations retaining chloride dependence of the respective wild type (WT). An osteopetrosis mutation of ClC-7 displaying fast gating kinetics (R762Q) was less sensitive to extracellular chloride compared to WT. The opposing substrate dependences of ClC-6 and ClC-7 Cl / H exchangers point to non-overlapping physiological functions, leading us to propose that inhibition of ClC-7 by luminal chloride and protons serves to prevent osmotic stress imposed by hyper-accumulation of chloride.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10842065PMC
http://dx.doi.org/10.1113/JP285431DOI Listing

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