Functional Analysis and Molecular Dynamics Simulation of LOX-1 K167N Polymorphism Reveal Alteration of Receptor Activity
Figure 2
In vivo interaction between wt-LOX-1 and mutated LOX-1 receptors.
(A) COS cells were co-transfected with wt-LOX-1-myc and wt-LOX-1-V5 (lanes 1 and 4), N/N167 LOX-1-V5 and wt-LOX-1-myc (lanes 2 and 5) and Sec-8H4-myc and wt-LOX-1-V5 (lanes 3 and 6) at a DNA ratio 1∶1. Half lysates were immunoblotted with anti-myc antibodies (lanes 1–3) and the remaining extracted proteins were first immunopurified with anti-V5 antibodies and then probed with anti-myc 9E10 (lanes 4–6). The migration of molecular weight markers (kDa) is indicated on the left and the heavy (γ) and the light (κ) chains are indicated on the right. (B) Lysates of COS cells co-expressing wt-LOX-1-myc and wt-LOX-1-V5 (lane 1 and 3) and wt-LOX-1-myc and N/N167 LOX-1-V5 (lanes 2 and 4) were separated by a NuPAGE 3–8% polyacrylamide gradient gel in non reducing conditions. Lanes 1 and 2 represent the extracts immunoblotted with Mab anti-V5, while lanes 3 and 4 are the extracted proteins first immunopurified with Rabbit anti-myc and then probed with Mab anti-V5. The dimers and tetramers are indicated, respectively, with D and T and the migration of molecular weight markers (kDa) is indicated on the left. All experiments were repeated three times with similar results.