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. 2009 Jul 27;186(2):201–209. doi: 10.1083/jcb.200904073

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© 2009 Brailoiu et al.

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Figure 5. — Mutation of a single residue in the putative pore region inactivates TPC1. (A) Multiple sequence alignment of the two putative pore regions (P1 and P2) of several animal TPCs. Asterisks highlight residues conserved in both pores of all isoforms from different species. Predicted helical region is outlined by the cartoon. (B and C) Cytosolic calcium responses of individual fura-2–loaded SKBR3 cells microinjected with either 10 nM (B) or 10 µM (C) NAADP. Cells were from mock-transfected cultures or cultures expressing TPC1 mutated in the putative pore region (TPC1 L273P). (D) Pooled data quantifying the magnitude of cytosolic changes under the various experimental conditions are shown. Error bars indicate SEM.