LdMT and LdRos3 form a stable complex in the <i>Leishmania</i> membrane.

<p>(A, B) Native- and SDS-PAGE analysis of LdMT and LdRos3-GFP. LdRos3-GFP was expressed in ΔLdMT parasites (A) and in ΔLdRos3 parasites (B). Solubilised membrane proteins were separated by native PAGE and analyzed for GFP fluorescence. The membrane extract obtained from ΔLdRos3 parasites contains a prominent fast-migrating fluorescent band (band I). The extract obtained from ΔLdMT parasites also contains slow-migrating fluorescent protein bands (band II and III). Regions of the gel corresponding to the fluorescent bands were excised and loaded onto SDS-PAGE gels which were subsequently immunoblotted with polyclonal antibodies against LdRos3 (α-LdRos3) and LdMT (α-LdMT). Size markers indicate relative mobility of proteins in kDa. (C) Immunoblots from co-immunoprecipitation assays. LdMT-GFP was immunoprecipated from a detergent-solubilised membrane fraction (load) obtained from ΔLdMT parasites expressing LdMT-GFP as well as non-transfected wild-type parasites (wt) using anti-GFP-MicroBeads. Immunoprecipitates (IP) were subjected to immunoblot analysis using antibodies that recognize LdMT (α-LdMT), LdRos3 (α-LdRos3) and metalloprotease gp63 (α-gp63).