The binding ability analysis of the normal VLDL receptor and its mutant.
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.
Qu
S
S
Department of Biochemistry, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030.
Feng
N
N
Liu
Z
Z
Zhou
H
H
Deng
Y
Y
Feng
Z
Z
D000818:Animals; D001665:Binding Sites; D016466:CHO Cells / Q000378:metabolism; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D008024:Ligands; D009154:Mutation; D009206:Myocardium / Q000378:metabolism*; D011973:Receptors, LDL / Q000096:biosynthesis / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014162:Transfection
D018076:DNA, Complementary; D008024:Ligands; D011973:Receptors, LDL; D011994:Recombinant Proteins; C036053:VLDL receptor