+ Site Statistics
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on Google+Follow on Google+
Follow on LinkedInFollow on LinkedIn

+ Translate

The low density lipoprotein receptor on human peripheral blood monocytes and lymphocytes: visualization by ligand blotting and immunoblotting techniques

Journal of Clinical Endocrinology and Metabolism 62(6): 1279-1287

The low density lipoprotein receptor on human peripheral blood monocytes and lymphocytes: visualization by ligand blotting and immunoblotting techniques

Western blotting and immunoprecipitation techniques were used to study low density lipoprotein (LDL) receptors from cultured human monocytes, lymphocytes, and fibroblasts. After incubation in lipoprotein-deficient media to allow induction, receptors were solubilized, subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to nitrocellulose paper, and detected by incubation with apolipoprotein B-containing lipoproteins followed by radiolabeled antiapolipoprotein B antibody. LDL receptors of identical apparent mol wt were demonstrated in monocyte and lymphocyte cell extracts; the receptors bound LDL as well as human post-prandial lipoprotein (density, less than 1.0063) on Western blots, but did not bind acetyl-LDL. LDL receptors in mononuclear cells could also be detected by immunoblotting using immunoglobulin G-C7, a monoclonal antibody raised against the bovine LDL receptor. When cell extracts were blotted, the mononuclear cell receptors had a lower mol wt than the fibroblast receptor in unreduced sodium dodecyl sulfate-polyacrylamide gels, with an apparent mol wt difference of 5,000 [134,000 +/- 1,400 (+/- SE) for mononuclear cells vs. 139,000 +/- 1600 for fibroblasts]. Immunoprecipitation and electrophoresis of L-[35S]methionine-labeled cell extracts revealed more rapid conversion of the receptor precursor to the mature receptor in monocytes than in fibroblasts. Western blotting of mononuclear cells from a patient with an abnormally high mol wt receptor characterized in fibroblasts demonstrated that the same mutation was expressed in monocytes and lymphocytes. This report represents the first visualization of human mononuclear cell LDL receptors by Western blotting and immunoprecipitation. These techniques may find application in population screening for LDL receptor variability.

(PDF same-day service: $19.90)

Accession: 006712210

PMID: 3700586

DOI: 10.1210/jcem-62-6-1279

Related references

Immunoblotting and ligand blotting of the low-density lipoprotein receptor of human liver, HepG2 cells and HeLa cells. Febs Letters 216(2): 275-280, 1987

Detection and quantitation of low density lipoprotein (LDL) receptors in human liver by ligand blotting, immunoblotting, and radioimmunoassay. LDL receptor protein content is correlated with plasma LDL cholesterol concentration. Journal of Biological Chemistry 261(36): 17127-17133, 1986

Visualization of the Chicken Oocyte Lipoprotein Receptor by Ligand Blotting with Biotinylated Plasma and Yolk Very Low Density Lipoproteins. Poultry Science 73(7): 1127-1136, 1994

Binding of low density lipoprotein and chylomicron remnants to the hepatic low density lipoprotein receptor of dogs rats and rabbits demonstrated by ligand blotting failure to detect a distinct chylomicron remnant binding protein by ligand blotting. European Journal of Biochemistry 159(2): 333-340, 1986

Ligand blotting visualization of the low density lipoprotein receptor in plasma membranes and in 2 classes of coated vesicles from adrenocortical cells. Febs Letters 184(2): 263-267, 1985

Low density lipoprotein receptor activity in freshly isolated human blood monocytes and lymphocytes. Metabolism: Clinical and Experimental 31(7): 721-727, 1982

Detection of the low-density-lipoprotein receptor with biotin-low-density lipoprotein. A rapid new method for ligand blotting. Biochemical Journal 229(3): 785-790, 1985

Identification of receptors for low density lipoprotein and acetylated low density lipoprotein in human placental microvilli by ligand immunoblotting. Biological Chemistry Hoppe-Seyler 366(9): 838, 1985

Low density lipoprotein receptor determination in peripheral blood mononuclear cells iii cell membrane constituents bind to low density lipoprotein and interfere with low density lipoprotein receptor analyses. 1985

Identification of the high density lipoprotein receptor in cultured cells by ligand blotting. American Heart Association Monograph (124): II-196, 1986