Evidence for the presence of lipid-free monomolecular apolipoprotein A-1 in plasma[S]
The first step in reverse cholesterol transport is a process by which lipid-free or lipid-poor apoA-1 removes cholesterol from cells through the action of ATP binding cassette transporter A1 at the plasma membrane. However the structure and composition of lipid-free or -poor apoA-1 in plasma remains...
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| Format: | Article |
| Language: | English |
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Elsevier
2014-02-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S002222752035029X |
| _version_ | 1819077233706270720 |
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| author | Osamu Miyazaki Jun Ogihara Isamu Fukamachi Takafumi Kasumi |
| author_facet | Osamu Miyazaki Jun Ogihara Isamu Fukamachi Takafumi Kasumi |
| author_sort | Osamu Miyazaki |
| collection | DOAJ |
| description | The first step in reverse cholesterol transport is a process by which lipid-free or lipid-poor apoA-1 removes cholesterol from cells through the action of ATP binding cassette transporter A1 at the plasma membrane. However the structure and composition of lipid-free or -poor apoA-1 in plasma remains obscure. We previously obtained a monoclonal antibody (MAb) that specifically recognizes apoA-1 in preβ1-HDL, the smallest apoA-1-containing particle in plasma, which we used to establish a preβ1-HDL ELISA. Here, we purified preβ1-HDL from fresh normal plasma using said antibody, and analyzed the composition and structure. ApoA-1 was detected, but neither phospholipid nor cholesterol were detected in the purified preβ1-HDL. Only globular, not discoidal, particles were observed by electron microscopy. In nondenaturing PAGE, no difference in the mobility was observed between the purified preβ1-HDL and original plasma preβ1-HDL, or between the preβ1-HDL and lipid-free apoA-1 prepared by delipidating HDL. In sandwich ELISA using two anti-preβ1-HDL MAbs, reactivity with intact plasma preβ1-HDL was observed in ELISA using two MAbs with distinct epitopes but no reactivity was observed in ELISA using a single MAb, and the same phenomenon was observed with monomolecular lipid-free apoA-1. These results suggest that plasma preβ1-HDL is lipid-free monomolecular apoA-1. |
| first_indexed | 2024-12-21T18:53:57Z |
| format | Article |
| id | doaj.art-52ede21aa9054afe92de9fbd7598733c |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-21T18:53:57Z |
| publishDate | 2014-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-52ede21aa9054afe92de9fbd7598733c2022-12-21T18:53:40ZengElsevierJournal of Lipid Research0022-22752014-02-01552214225Evidence for the presence of lipid-free monomolecular apolipoprotein A-1 in plasma[S]Osamu Miyazaki0Jun Ogihara1Isamu Fukamachi2Takafumi Kasumi3To whom correspondence should be addressed; Tsukuba Research Institute, Research and Development Division, Sekisui Medical Co., Ltd., Ibaraki 301-0852, JapanDepartment of Biological Chemistry, College of Biosource Sciences, Nihon University, Kanagawa 252-0880, JapanTsukuba Research Institute, Research and Development Division, Sekisui Medical Co., Ltd., Ibaraki 301-0852, JapanDepartment of Biological Chemistry, College of Biosource Sciences, Nihon University, Kanagawa 252-0880, JapanThe first step in reverse cholesterol transport is a process by which lipid-free or lipid-poor apoA-1 removes cholesterol from cells through the action of ATP binding cassette transporter A1 at the plasma membrane. However the structure and composition of lipid-free or -poor apoA-1 in plasma remains obscure. We previously obtained a monoclonal antibody (MAb) that specifically recognizes apoA-1 in preβ1-HDL, the smallest apoA-1-containing particle in plasma, which we used to establish a preβ1-HDL ELISA. Here, we purified preβ1-HDL from fresh normal plasma using said antibody, and analyzed the composition and structure. ApoA-1 was detected, but neither phospholipid nor cholesterol were detected in the purified preβ1-HDL. Only globular, not discoidal, particles were observed by electron microscopy. In nondenaturing PAGE, no difference in the mobility was observed between the purified preβ1-HDL and original plasma preβ1-HDL, or between the preβ1-HDL and lipid-free apoA-1 prepared by delipidating HDL. In sandwich ELISA using two anti-preβ1-HDL MAbs, reactivity with intact plasma preβ1-HDL was observed in ELISA using two MAbs with distinct epitopes but no reactivity was observed in ELISA using a single MAb, and the same phenomenon was observed with monomolecular lipid-free apoA-1. These results suggest that plasma preβ1-HDL is lipid-free monomolecular apoA-1.http://www.sciencedirect.com/science/article/pii/S002222752035029Xpre-beta-high density lipoproteinlipid-free apolipoprotein A-1reverse cholesterol transport |
| spellingShingle | Osamu Miyazaki Jun Ogihara Isamu Fukamachi Takafumi Kasumi Evidence for the presence of lipid-free monomolecular apolipoprotein A-1 in plasma[S] Journal of Lipid Research pre-beta-high density lipoprotein lipid-free apolipoprotein A-1 reverse cholesterol transport |
| title | Evidence for the presence of lipid-free monomolecular apolipoprotein A-1 in plasma[S] |
| title_full | Evidence for the presence of lipid-free monomolecular apolipoprotein A-1 in plasma[S] |
| title_fullStr | Evidence for the presence of lipid-free monomolecular apolipoprotein A-1 in plasma[S] |
| title_full_unstemmed | Evidence for the presence of lipid-free monomolecular apolipoprotein A-1 in plasma[S] |
| title_short | Evidence for the presence of lipid-free monomolecular apolipoprotein A-1 in plasma[S] |
| title_sort | evidence for the presence of lipid free monomolecular apolipoprotein a 1 in plasma s |
| topic | pre-beta-high density lipoprotein lipid-free apolipoprotein A-1 reverse cholesterol transport |
| url | http://www.sciencedirect.com/science/article/pii/S002222752035029X |
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