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In a previous study, it was shown that the phosphatidylinositol-linked plasma membrane protein, gp60, is the low-density lipoprotein (LDL) receptor of rat alveolar epithelial cells (AEC) in vitro. Here we demonstrate, using a combination of molecular biological, biophysical, and immunocytochemical techniques, that gp60 functions as the high-density lipoprotein (HDL) receptor of AEC in this culture system. Scatchard analysis of 125I-HDL binding to AEC at 4 degrees C revealed a single class of binding sites with a Kd of 1.4 x 10(-8) M and a binding capacity of 0.8 pmol/mg of cell protein. Competition studies indicated that the HDL-receptor complex was functionally active at 4 degrees C, suggesting that it was a functional lipid-protein complex. These studies were confirmed and extended by demonstrating that a mutant form of gp60, expressed in COS cells, exhibited specific and saturable HDL binding. Affinity labeling of gp60 by 125I-HDL-HSA was followed by SDS-gel electrophoresis, demonstrating a molecular mass of about 60 kDa. The partially purified receptor was susceptible to degradation by N-glycanase and O-glycanase, indicating that part of the gp60 molecule consists of N- and O-glycans. Furthermore, in Western blot analysis of rat lung plasma membrane proteins, a receptor-like protein was detected with an apparent molecular mass of 60 kDa, which cross-reacted with anti-HDL receptor antibody. These results indicate that gp60 is the HDL receptor of AEC and that it functions as a receptor in its native membrane environment.Video: The Amazing Evolution of Ashley Judd

In this week’s video (below), Ashley Judd explains how life lessons influence her work as an actress.

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