Antipeptide antibody against the human low-density-lipoprotein receptor. Characterization and cross-reactivity with bovine lymphocytes.
A dodecapeptide corresponding to the external N-terminal sequence of the human low-density-lipoprotein (LDL) receptor was synthesized. Antibodies raised in rabbits against the peptide were purified and were shown to react specifically with the peptide and with human LDL receptor of fibroblasts, HeLa cells and lymphocytes using binding studies and immunoblotting. By indirect immunogold analysis, antibodies bound to the LDL receptor of human lymphocytes could be revealed as clusters. Anti-receptor peptide immunoglobulins specifically bound to the human HeLa cell's LDL receptor with a lower affinity than LDL (Kd x 3). The anti-receptor peptide immunoglobulins and 125I-labelled-LDL competed with each other for the LDL-receptor sites. Antibodies failed to react with lymphocytes of subjects with the homozygous form of familial hypercholesterolaemia. Cross-reactivity with the dodecapeptide of the bovine LDL receptor was limited, but this cross-reactivity was confirmed by the binding of anti-receptor peptide immunoglobulins to the LDL receptor from bovine lymphocytes.
Agnani
G
G
SERLIA et INSERM U 279 Institut Pasteur, Lille, France.
Delpierre
C
C
Benhamamouch
S
S
Torpier
G
G
Cachera
C
C
Koffigan
M
M
Tartar
A
A
Fruchart
J C
JC
Clavey
V
V
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D001667:Binding, Competitive; D015153:Blotting, Western; D002417:Cattle; D003429:Cross Reactions; D004705:Endocytosis; D005347:Fibroblasts / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells / Q000276:immunology; D006801:Humans; D007150:Immunohistochemistry; D008077:Lipoproteins, LDL / Q000378:metabolism; D008214:Lymphocytes / Q000276:immunology; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D011973:Receptors, LDL / Q000276:immunology* / Q000378:metabolism
D008077:Lipoproteins, LDL; D010455:Peptides; D011973:Receptors, LDL