Isolation of allele-specific, receptor-binding-defective low density lipoproteins from familial defective apolipoprotein B-100 subjects.
Familial defective apolipoprotein B-100 (FDB) is a genetic disorder apparently caused by a single amino acid substitution (Arg3500–>Gln) that disrupts the binding of low density lipoproteins (LDL) to the LDL receptor. The plasma of FDB heterozygotes contains a mixture of normal LDL and LDL that i...
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1994-08-01
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Series: | Journal of Lipid Research |
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doaj-b80692fa64d2496c8a5b1a82a01aefe92021-04-26T05:51:06ZengElsevierJournal of Lipid Research0022-22751994-08-0135814691476Isolation of allele-specific, receptor-binding-defective low density lipoproteins from familial defective apolipoprotein B-100 subjects.K S Arnold0M E Balestra1R M Krauss2L K Curtiss3S G Young4T L Innerarity5Gladstone Institute of Cardiovascular Disease, University of California, San Francisco 94141-9100.Gladstone Institute of Cardiovascular Disease, University of California, San Francisco 94141-9100.Gladstone Institute of Cardiovascular Disease, University of California, San Francisco 94141-9100.Gladstone Institute of Cardiovascular Disease, University of California, San Francisco 94141-9100.Gladstone Institute of Cardiovascular Disease, University of California, San Francisco 94141-9100.Gladstone Institute of Cardiovascular Disease, University of California, San Francisco 94141-9100.Familial defective apolipoprotein B-100 (FDB) is a genetic disorder apparently caused by a single amino acid substitution (Arg3500–>Gln) that disrupts the binding of low density lipoproteins (LDL) to the LDL receptor. The plasma of FDB heterozygotes contains a mixture of normal LDL and LDL that is defective in binding to the LDL receptor. In this study, the monoclonal antibody MB19 (which recognizes an immunogenetic polymorphism in apolipoprotein B-100) was used to determine the percentage of defective LDL in the plasma of FDB heterozygotes and to isolate allele-specific receptor-binding-defective LDL. Several FDB heterozygotes were identified who were heterozygous for the MB19 polymorphism: one apolipoprotein B allotype in each of these individuals bound with low affinity to MB19 and possessed the Arg3500–>Gln mutation, whereas the other apolipoprotein B allotype bound with high affinity to MB19 and normally to the LDL receptor. Using MB19 radio-immunoassay, we determined that an average of 73% (range 65-87) of the total LDL from FDB heterozygotes contained the Arg3500–>Gln mutation. Antibody MB19-Sepharose immuno-affinity chromatography was used to separate the receptor-binding-defective LDL from the normal LDL. The isolated LDL contained primarily the Arg3500–>Gln mutation and had only about 9% of normal LDL receptor-binding ability. Finally, the MB19-Sepharose chromatography procedure may be useful for isolating other allele-specific LDL that have functionally significant mutations.http://www.sciencedirect.com/science/article/pii/S0022227520400884 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
K S Arnold M E Balestra R M Krauss L K Curtiss S G Young T L Innerarity |
spellingShingle |
K S Arnold M E Balestra R M Krauss L K Curtiss S G Young T L Innerarity Isolation of allele-specific, receptor-binding-defective low density lipoproteins from familial defective apolipoprotein B-100 subjects. Journal of Lipid Research |
author_facet |
K S Arnold M E Balestra R M Krauss L K Curtiss S G Young T L Innerarity |
author_sort |
K S Arnold |
title |
Isolation of allele-specific, receptor-binding-defective low density lipoproteins from familial defective apolipoprotein B-100 subjects. |
title_short |
Isolation of allele-specific, receptor-binding-defective low density lipoproteins from familial defective apolipoprotein B-100 subjects. |
title_full |
Isolation of allele-specific, receptor-binding-defective low density lipoproteins from familial defective apolipoprotein B-100 subjects. |
title_fullStr |
Isolation of allele-specific, receptor-binding-defective low density lipoproteins from familial defective apolipoprotein B-100 subjects. |
title_full_unstemmed |
Isolation of allele-specific, receptor-binding-defective low density lipoproteins from familial defective apolipoprotein B-100 subjects. |
title_sort |
isolation of allele-specific, receptor-binding-defective low density lipoproteins from familial defective apolipoprotein b-100 subjects. |
publisher |
Elsevier |
series |
Journal of Lipid Research |
issn |
0022-2275 |
publishDate |
1994-08-01 |
description |
Familial defective apolipoprotein B-100 (FDB) is a genetic disorder apparently caused by a single amino acid substitution (Arg3500–>Gln) that disrupts the binding of low density lipoproteins (LDL) to the LDL receptor. The plasma of FDB heterozygotes contains a mixture of normal LDL and LDL that is defective in binding to the LDL receptor. In this study, the monoclonal antibody MB19 (which recognizes an immunogenetic polymorphism in apolipoprotein B-100) was used to determine the percentage of defective LDL in the plasma of FDB heterozygotes and to isolate allele-specific receptor-binding-defective LDL. Several FDB heterozygotes were identified who were heterozygous for the MB19 polymorphism: one apolipoprotein B allotype in each of these individuals bound with low affinity to MB19 and possessed the Arg3500–>Gln mutation, whereas the other apolipoprotein B allotype bound with high affinity to MB19 and normally to the LDL receptor. Using MB19 radio-immunoassay, we determined that an average of 73% (range 65-87) of the total LDL from FDB heterozygotes contained the Arg3500–>Gln mutation. Antibody MB19-Sepharose immuno-affinity chromatography was used to separate the receptor-binding-defective LDL from the normal LDL. The isolated LDL contained primarily the Arg3500–>Gln mutation and had only about 9% of normal LDL receptor-binding ability. Finally, the MB19-Sepharose chromatography procedure may be useful for isolating other allele-specific LDL that have functionally significant mutations. |
url |
http://www.sciencedirect.com/science/article/pii/S0022227520400884 |
work_keys_str_mv |
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