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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.
For over 20 years, Judd (Actress, The Cotton Club, The_Hun
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