Figure 5. Intracellular distribution of ATP7B in COS-1 cells expressing WT ATP7B (A) or ATP7B subjected to mutations at Asp¹⁰²⁷ (B), at Ser⁴⁷⁸, Ser⁴⁸¹, Ser¹¹²¹ and Ser¹⁴⁵³ (C), at the transmembrane domain (TMBS) copper site (D) or at the sixth NMBD copper site (E).

Note the presence of cytosolic trafficking vesicles of WT ATP7B and even following Asp¹⁰²⁷ mutation, but no trafficking following serine, NMBD or TMBS copper site mutations. All panels present different fields of cells treated identically. A copper load (200 μM) was added to the culture medium 2 h following infection with adenovirus vector for delivery of ATP7B cDNA (WT or mutant). The cells were fixed 24 h later. Secondary antibodies were goat anti-mouse conjugated with Alexa Fluor® 488 for the ATP7B c-Myc tag (green) and donkey anti-rabbit conjugated with Cy5 (indodicarbocyanine) for Golgi (blue). Red indicates nuclei stained with propidium iodide. Scale bar, 10 μm. This Figure was adapted from J. Biol. Chem. [60] Pilankatta, R., Lewis, D. and Inesi G. Involvement of protein kinase D in expression and trafficking of ATP7B (copper ATPase). Journal of Biological Chemistry. 2011; 286, 7389–7396.