Purification of the low density lipoprotein receptor, an acidic glycoprotein of 164,000 molecular weight
- PMID: 6277909
Purification of the low density lipoprotein receptor, an acidic glycoprotein of 164,000 molecular weight
Abstract
This paper describes a rapid two-step procedure for the purification of the low density lipoprotein receptor from bovine adrenal cortex membranes. After solubilization with nonionic detergents, the receptor adheres tightly to a DEAE-cellulose column at pH 6. Following elution from DEAE-cellulose, detergent is removed, leaving the receptor in a soluble form. The receptor is then subjected to affinity chromatography on low density lipoprotein coupled to Sepharose 4B. The receptor is eluted with suramin, a newly-found inhibitor of low density lipoprotein-receptor interactions. This procedure yields a single protein with a molecular weight of 164,000. The same protein is also isolated when the crude DEAE-cellulose fraction is applied to an immunoaffinity column containing a monoclonal antibody directed against the receptor. The 164,000-dalton receptor protein has an acidic isoelectric point of 4.6, which rises to 4.8 after extensive treatment with neuraminidase. The purified receptor retains all of the binding properties of the receptor of intact cells and crude membranes.
Similar articles
-
Partial purification and characterization of the low density lipoprotein receptor from bovine adrenal cortex.J Biol Chem. 1980 Dec 10;255(23):11442-7. J Biol Chem. 1980. PMID: 6254970
-
Reconstitution of the low density lipoprotein receptor.J Cell Biochem. 1983;23(1-4):95-106. doi: 10.1002/jcb.240230110. J Cell Biochem. 1983. PMID: 6327742
-
Isolation and characterization of the apolipoprotein E receptor from canine and human liver.J Biol Chem. 1986 Mar 25;261(9):4256-67. J Biol Chem. 1986. PMID: 3005328
-
Isolation and characterization of a lipoprotein receptor from the fat body of an insect, Manduca sexta.J Biol Chem. 1990 Apr 5;265(10):5761-7. J Biol Chem. 1990. PMID: 2156827
-
Visualization of lipoprotein receptors by ligand blotting.J Biol Chem. 1983 Apr 10;258(7):4606-11. J Biol Chem. 1983. PMID: 6300091
Cited by
-
Characterization of lipophorin receptor (LpR) mediating the binding of high density lipophorin (HDLp) in the silkworm, Bombyx mori.J Insect Sci. 2011;11:150. doi: 10.1673/031.011.15001. J Insect Sci. 2011. PMID: 22236107 Free PMC article.
-
Immunoassay of bovine and human low-density-lipoprotein receptors using monoclonal antibodies.Biochem J. 1986 Sep 1;238(2):405-10. doi: 10.1042/bj2380405. Biochem J. 1986. PMID: 3800945 Free PMC article.
-
Specific postendocytic proteolysis of apolipoprotein B in oocytes does not abolish receptor recognition.Proc Natl Acad Sci U S A. 1989 Feb;86(3):906-10. doi: 10.1073/pnas.86.3.906. Proc Natl Acad Sci U S A. 1989. PMID: 2644651 Free PMC article.
-
The human alpha 2-macroglobulin receptor: identification of a 420-kD cell surface glycoprotein specific for the activated conformation of alpha 2-macroglobulin.J Cell Biol. 1990 Apr;110(4):1041-8. doi: 10.1083/jcb.110.4.1041. J Cell Biol. 1990. PMID: 1691187 Free PMC article.
-
Detection of the low-density-lipoprotein receptor with biotin-low-density lipoprotein. A rapid new method for ligand blotting.Biochem J. 1985 Aug 1;229(3):785-90. doi: 10.1042/bj2290785. Biochem J. 1985. PMID: 4052026 Free PMC article.
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources
