Direct detection of ABCA1-dependent HDL formation based on lipidation-induced hydrophobicity change in apoA-I[S]
ABCA1 mediates the efflux of cholesterol and phospholipids into apoA-I to form HDL, which is important in the prevention of atherosclerosis. To develop a novel method for the evaluation of HDL formation, we prepared an apoA-I-POLARIC by labeling the specific residue of an apoA-I variant with a hydro...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2014-11-01
|
| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520349944 |
| _version_ | 1818381583133966336 |
|---|---|
| author | Risa Omura Kohjiro Nagao Norihiro Kobayashi Kazumitsu Ueda Hiroyuki Saito |
| author_facet | Risa Omura Kohjiro Nagao Norihiro Kobayashi Kazumitsu Ueda Hiroyuki Saito |
| author_sort | Risa Omura |
| collection | DOAJ |
| description | ABCA1 mediates the efflux of cholesterol and phospholipids into apoA-I to form HDL, which is important in the prevention of atherosclerosis. To develop a novel method for the evaluation of HDL formation, we prepared an apoA-I-POLARIC by labeling the specific residue of an apoA-I variant with a hydrophobicity-sensitive fluorescence probe that detects the environmental change around apoA-I during HDL formation. apoA-I-POLARIC possesses the intact ABCA1-dependent HDL formation activity and shows 4.0-fold higher fluorescence intensity in HDL particles than in the lipid-free state. Incubation of apoA-I-POLARIC with ABCA1-expressing cells, but not ABCA1-non-expressing cells, caused a 1.7-fold increase in fluorescence intensity. Gel filtration analysis demonstrated that the increase in fluorescence intensity of apoA-I-POLARIC represents the amount of apoA-I incorporated into the discoidal HDL particles rather than the amount of secreted cholesterol. THP-1 macrophage-mediated HDL formation and inhibition of HDL formation by cyclosporine A could also be measured using apoA-I-POLARIC. Furthermore, HDL formation-independent lipid release induced by microparticle formation or cell death was not detected by apoA-I-POLARIC. These results demonstrate that HDL formation by ABCA1-expressing cells can be specifically detected by sensing hydrophobicity change in apoA-I, thus providing a novel method for assessing HDL formation and screening of the HDL formation modulator. |
| first_indexed | 2024-12-14T02:36:53Z |
| format | Article |
| id | doaj.art-00346a7bf5b245e08379514c04f56423 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-14T02:36:53Z |
| publishDate | 2014-11-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-00346a7bf5b245e08379514c04f564232022-12-21T23:20:06ZengElsevierJournal of Lipid Research0022-22752014-11-01551124232431Direct detection of ABCA1-dependent HDL formation based on lipidation-induced hydrophobicity change in apoA-I[S]Risa Omura0Kohjiro Nagao1Norihiro Kobayashi2Kazumitsu Ueda3Hiroyuki Saito4Institute of Health Biosciences and Graduate School of Pharmaceutical Sciences, The University of Tokushima, Tokushima 770-8505, JapanInstitute of Health Biosciences and Graduate School of Pharmaceutical Sciences, The University of Tokushima, Tokushima 770-8505, Japan; To whom correspondence should be addressedKobe Pharmaceutical University, Kobe 658-8558, JapanLaboratory of Cellular Biochemistry, Division of Applied Life Sciences, Kyoto University Graduate School of Agriculture, Kyoto 606-8502, Japan; Institute for Integrated Cell-Material Sciences (iCeMS), Kyoto University, Kyoto 606-8502, JapanInstitute of Health Biosciences and Graduate School of Pharmaceutical Sciences, The University of Tokushima, Tokushima 770-8505, JapanABCA1 mediates the efflux of cholesterol and phospholipids into apoA-I to form HDL, which is important in the prevention of atherosclerosis. To develop a novel method for the evaluation of HDL formation, we prepared an apoA-I-POLARIC by labeling the specific residue of an apoA-I variant with a hydrophobicity-sensitive fluorescence probe that detects the environmental change around apoA-I during HDL formation. apoA-I-POLARIC possesses the intact ABCA1-dependent HDL formation activity and shows 4.0-fold higher fluorescence intensity in HDL particles than in the lipid-free state. Incubation of apoA-I-POLARIC with ABCA1-expressing cells, but not ABCA1-non-expressing cells, caused a 1.7-fold increase in fluorescence intensity. Gel filtration analysis demonstrated that the increase in fluorescence intensity of apoA-I-POLARIC represents the amount of apoA-I incorporated into the discoidal HDL particles rather than the amount of secreted cholesterol. THP-1 macrophage-mediated HDL formation and inhibition of HDL formation by cyclosporine A could also be measured using apoA-I-POLARIC. Furthermore, HDL formation-independent lipid release induced by microparticle formation or cell death was not detected by apoA-I-POLARIC. These results demonstrate that HDL formation by ABCA1-expressing cells can be specifically detected by sensing hydrophobicity change in apoA-I, thus providing a novel method for assessing HDL formation and screening of the HDL formation modulator.http://www.sciencedirect.com/science/article/pii/S0022227520349944cholesterol effluxPOLARIChydrophobicity-sensitive fluorescence probeATP binding cassette transporter A1high density lipoproteinapolipoprotein A-I |
| spellingShingle | Risa Omura Kohjiro Nagao Norihiro Kobayashi Kazumitsu Ueda Hiroyuki Saito Direct detection of ABCA1-dependent HDL formation based on lipidation-induced hydrophobicity change in apoA-I[S] Journal of Lipid Research cholesterol efflux POLARIC hydrophobicity-sensitive fluorescence probe ATP binding cassette transporter A1 high density lipoprotein apolipoprotein A-I |
| title | Direct detection of ABCA1-dependent HDL formation based on lipidation-induced hydrophobicity change in apoA-I[S] |
| title_full | Direct detection of ABCA1-dependent HDL formation based on lipidation-induced hydrophobicity change in apoA-I[S] |
| title_fullStr | Direct detection of ABCA1-dependent HDL formation based on lipidation-induced hydrophobicity change in apoA-I[S] |
| title_full_unstemmed | Direct detection of ABCA1-dependent HDL formation based on lipidation-induced hydrophobicity change in apoA-I[S] |
| title_short | Direct detection of ABCA1-dependent HDL formation based on lipidation-induced hydrophobicity change in apoA-I[S] |
| title_sort | direct detection of abca1 dependent hdl formation based on lipidation induced hydrophobicity change in apoa i s |
| topic | cholesterol efflux POLARIC hydrophobicity-sensitive fluorescence probe ATP binding cassette transporter A1 high density lipoprotein apolipoprotein A-I |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520349944 |
| work_keys_str_mv | AT risaomura directdetectionofabca1dependenthdlformationbasedonlipidationinducedhydrophobicitychangeinapoais AT kohjironagao directdetectionofabca1dependenthdlformationbasedonlipidationinducedhydrophobicitychangeinapoais AT norihirokobayashi directdetectionofabca1dependenthdlformationbasedonlipidationinducedhydrophobicitychangeinapoais AT kazumitsuueda directdetectionofabca1dependenthdlformationbasedonlipidationinducedhydrophobicitychangeinapoais AT hiroyukisaito directdetectionofabca1dependenthdlformationbasedonlipidationinducedhydrophobicitychangeinapoais |