The binding ability analysis of the normal VLDL receptor and its mutant

J Tongji Med Univ. 2001;21(3):177-80, 194. doi: 10.1007/BF02886422.

Abstract

The ligand-binding domain of VLDL receptor contains eight imperfectly similar repeats. To discuss the contribution of each repeat to ligand binding, the RT-PCR technique was used to clone the VLDLR-cDNA from the heart muscle of Chinese people. Two recombinants were further constructed, which contained the full-length cDNA of VLDLR and the mutant lacking repeats 1-5. CHO cell line was transfected with two recombinants. The expression of VLDLR gene could be detected by RT-PCR from the CHO cells transfected with pCD-VR. The results of binding experiments showed that the ability of the CHO cells transfected with the full-length cDNA of VLDL-R binding DiI-labeled beta-VLDL was higher than that of the CHO cells transfected with the mutant. Our findings indicated that human VLDL-R gene could be expressed effectively on CHO cells, and the receptor was almost inactivated when repeats 1-5 were deleted.

MeSH terms

  • Animals
  • Binding Sites
  • CHO Cells / metabolism
  • Cloning, Molecular
  • Cricetinae
  • DNA, Complementary / genetics
  • Humans
  • Ligands
  • Mutation*
  • Myocardium / metabolism*
  • Receptors, LDL / biosynthesis
  • Receptors, LDL / genetics*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Transfection

Substances

  • DNA, Complementary
  • Ligands
  • Receptors, LDL
  • Recombinant Proteins
  • VLDL receptor