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

• Published in: Current medical science 2001, Vol.21 (3), p.177-180
• Main Authors: Qu, S, Feng, N, Liu, Z, Zhou, H, Deng, Y, Feng, Z
• Format: Article
• Language: English
• Subjects: 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
• ISSN: 0257-716X; 2096-5230; 2523-899X
• DOI: 10.1007/bf02886422

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.