Mapping of human apolipoprotein B antigenic determinants.

Marcel, Yves L.; Innerarity, Thomas L.; Spilman, C. H.; Mahley, Robert W.; Protter, Andrew A.; Milne, Ross W.

doi:10.1161/01.atv.7.2.166

Cited by 85 publications

(34 citation statements)

References 27 publications

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“…All of these residues lie on helix 4, one of five helices located in the N-terminal domain of apoE [21]. The regions of human apoB-100 that contain such clusters have been implicated in binding to the LDL receptor (residues 3359-3368, Arg-LeuThr-Arg-Lys-Arg-Gly-Leu-Lys-Leu) [22,23]. Furthermore it has been shown that two such clusters of VTG, residues 493498 (Leu-Lys-Arg-Ile-Leu-Lys) and residues 1079-1084 (Lys-Leu-Lys-Arg-Ile-Leu), mediate binding to the VLDL/ VTG receptor.…”

Section: Discussionmentioning

confidence: 99%

Very low density lipoprotein receptor binds apolipoprotein E2/2 as well as apolipoprotein E3/3

et al. 1996

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Section: Discussionmentioning

confidence: 99%

Very low density lipoprotein receptor binds apolipoprotein E2/2 as well as apolipoprotein E3/3

et al. 1996

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“…Peptides corresponding to amino acids 17-27, 104-119, and 204-217 of the bovine scavenger receptor 8 were synthesized at the Biomolecular Resource Center, University of California, San Francisco. To facilitate coupling to keyhole limpet hemocyanine 9 before production of antibodies in guinea pigs, all the synthesized peptides possessed an amino-terminal cysteine.…”

Section: Methodsmentioning

confidence: 99%

Further characterization of the acetyl LDL (scavenger) receptor expressed by rabbit smooth muscle cells and fibroblasts.

Pitas

Friera

McGuire

et al. 1992

Arterioscler Thromb

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We have previously demonstrated that the acetyl low density lipoprotein (LDL), or scavenger, receptor is expressed by rabbit smooth muscle cells (SMCs) and fibroblasts. Moreover, receptor activity in rabbit fibroblasts was regulated over a wide range by preincubating the cells with secretion products from human platelets or with phorbol esters. The current studies were undertaken to examine the regulation of receptor expression in rabbit SMCs, to characterize further the receptor expressed by rabbit SMCs and fibroblasts, and to determine whether incubating these cells with chemically modified lipoproteins would lead to foam cell formation. Receptor activity was increased fivefold in rabbit SMCs by preincubation with platelet secretion products and nine-to 20-fold by preincubation with phorbol esters or mezerein, a non-phorbol activator of protein kinase C. The phorbol ester-induced increase in receptor activity was due to an increased mass of scavenger receptor as determined by both ligand blots and immunoblots of membrane proteins from these cells. The immunoblotting studies suggest that the SMC scavenger receptor activity is due to type I or type II scavenger receptors.

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“…While they were useful for both of these applications [4,8,9], they also provide an interesting panel of Mabs to compare the structure of apoB in the presence or absence of lipids. As such, they complement the other panel of Mabs against LDL apoB [lo], and allow us to define which parts of apoB are exposed on the surface of LDL and which parts are not accessible.…”

mentioning

confidence: 99%

Primary sequence mapping of human apolipoprotein B‐100 epitopes

Chen

Marcel

Yang

et al. 1988

European Journal of Biochemistry

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Differential trypsin-accessibility and monoclonal antibodies (Mabs) to human apolipoprotein (apo) B-1 00 are both important tools for probing apoB structure and conformation on low-density lipoproteins (LDL). In this study, we have mapped > 80% of the C-terminal region (720 residues) of LDL apoB-100 using trypsin digestion.Our results extend our previous data Nature (Lond.) 323, confirming that the C-terminal region of about 420 residues of apoB-100 is largely inaccessible to trypsin, whereas the part just preceding this region has interspersed trypsin-accessible and inaccessible peptides. We have determined the amino acid sequence of specific apoB-100 peptides containing epitopes recognized by four separate Mabs: two epitopes have been mapped to within 20 residues, one has been mapped to 36 residues, and the last to 80 residues. We used polyclonal antisera to identify 16 overlapping clones of varying lengths of apoB-100 cDNAs extending from the C-terminus of apoB-100 cloned in the expression vector, Agtll . These clones were then tested against individual Mabs. By nucleotide sequence analysis of overlapping clones that show differential reactivities to different Mabs, we have mapped the individual epitopes of each Mab to within about 50 -150 amino acid residues predicted from the DNA sequences. Confirmation and further fine mapping were accomplished by competition for LDL binding using partially purified fusion proteins and chemically synthesized oligopeptides. Two epitopes (Mabs 7 and 22) were mapped to the C-terminal20 amino acids of apoB-100, one (Mab 16) to residues 4154-4189, and another (Mab 20) to residues 3926-4005. Mab 16 precipitates more than 80% of LDL particles. Mab 20 precipitates only denatured apoB but not native LDL apoB Mol. Zmmunol. 24, 4351. Mabs 7 and 22 are unique in that they precipitate LDL apoB modified by storage much better than freshly isolated LDLapoB. Although epitope expression and trypsin-accessibility represent two useful probes for the study of protein conformation, there was no obvious correlation between these two parameters when applied to LDL apoB for the antibodies we have examined Apolipoprotein (apo) B is the largest of the apolipoproteins. It is heterogeneous in size; apoB-100, a 512-kDa protein, is an obligatory component in very-low-density, intermediate-density, and low-density lipoproteins (LDL); apoB-48, a smaller protein of about 264 kDa, is present mainly on chylomicrons [l]. Because of the central role of apoB in lipid transport and in cholesterol homeostasis, it is important to understand the structure and conformation of apoB in the various classes of lipoprotein particles. The sequence analysis of cloned apoB-100 cDNAs has provided the complete primary sequence of the protein [2 -61. However, in order to understand fully the conformation of apoB-100 on the lipoprotein particles, additional molecular probes are needed to define the exposed vs the buried regions of the protein on these particles. One method is the trypsin digestion of apoB-I 00 on the lipop...

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Mapping of human apolipoprotein B antigenic determinants.

Cited by 85 publications

References 27 publications

Very low density lipoprotein receptor binds apolipoprotein E2/2 as well as apolipoprotein E3/3

Very low density lipoprotein receptor binds apolipoprotein E2/2 as well as apolipoprotein E3/3

Further characterization of the acetyl LDL (scavenger) receptor expressed by rabbit smooth muscle cells and fibroblasts.

Primary sequence mapping of human apolipoprotein B‐100 epitopes

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