ATP hydrolysis-dependent conformational changes in the extracellular domain of ABCA1 are associated with apoA-I binding[S]
ATP-binding cassette protein A1 (ABCA1) plays a major role in cholesterol homeostasis and high-density lipoprotein (HDL) metabolism. Although it is predicted that apolipoprotein A-I (apoA-I) directly binds to ABCA1, the physiological importance of this interaction is still controversial and the conf...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2012-01-01
|
| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520408065 |
| _version_ | 1818611340366839808 |
|---|---|
| author | Kohjiro Nagao Kei Takahashi Yuya Azuma Mie Takada Yasuhisa Kimura Michinori Matsuo Noriyuki Kioka Kazumitsu Ueda |
| author_facet | Kohjiro Nagao Kei Takahashi Yuya Azuma Mie Takada Yasuhisa Kimura Michinori Matsuo Noriyuki Kioka Kazumitsu Ueda |
| author_sort | Kohjiro Nagao |
| collection | DOAJ |
| description | ATP-binding cassette protein A1 (ABCA1) plays a major role in cholesterol homeostasis and high-density lipoprotein (HDL) metabolism. Although it is predicted that apolipoprotein A-I (apoA-I) directly binds to ABCA1, the physiological importance of this interaction is still controversial and the conformation required for apoA-I binding is unclear. In this study, the role of the two nucleotide-binding domains (NBD) of ABCA1 in apoA-I binding was determined by inserting a TEV protease recognition sequence in the linker region of ABCA1. Analyses of ATP binding and occlusion to wild-type ABCA1 and various NBD mutants revealed that ATP binds equally to both NBDs and is hydrolyzed at both NBDs. The interaction with apoA-I and the apoA-I-dependent cholesterol efflux required not only ATP binding but also hydrolysis in both NBDs. NBD mutations and cellular ATP depletion decreased the accessibility of antibodies to a hemagglutinin (HA) epitope that was inserted at position 443 in the extracellular domain (ECD), suggesting that the conformation of ECDs is altered by ATP hydrolysis at both NBDs. These results suggest that ATP hydrolysis at both NBDs induces conformational changes in the ECDs, which are associated with apoA-I binding and cholesterol efflux. |
| first_indexed | 2024-12-16T15:28:46Z |
| format | Article |
| id | doaj.art-f18d2aaec6ac41189a2989b8ede00ad8 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-16T15:28:46Z |
| publishDate | 2012-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-f18d2aaec6ac41189a2989b8ede00ad82022-12-21T22:26:25ZengElsevierJournal of Lipid Research0022-22752012-01-01531126136ATP hydrolysis-dependent conformational changes in the extracellular domain of ABCA1 are associated with apoA-I binding[S]Kohjiro Nagao0Kei Takahashi1Yuya Azuma2Mie Takada3Yasuhisa Kimura4Michinori Matsuo5Noriyuki Kioka6Kazumitsu Ueda7Institute for Integrated Cell-Material Sciences (iCeMS), Kyoto University, Kyoto 606-8502, JapanLaboratory of Cellular Biochemistry, Division of Applied Life Sciences, Graduate School of Agriculture andLaboratory of Cellular Biochemistry, Division of Applied Life Sciences, Graduate School of Agriculture andLaboratory of Cellular Biochemistry, Division of Applied Life Sciences, Graduate School of Agriculture andLaboratory of Cellular Biochemistry, Division of Applied Life Sciences, Graduate School of Agriculture andLaboratory of Cellular Biochemistry, Division of Applied Life Sciences, Graduate School of Agriculture andLaboratory of Cellular Biochemistry, Division of Applied Life Sciences, Graduate School of Agriculture andTo whom correspondence should be addressed; Institute for Integrated Cell-Material Sciences (iCeMS), Kyoto University, Kyoto 606-8502, Japan; Laboratory of Cellular Biochemistry, Division of Applied Life Sciences, Graduate School of Agriculture and; To whom correspondence should be addressedATP-binding cassette protein A1 (ABCA1) plays a major role in cholesterol homeostasis and high-density lipoprotein (HDL) metabolism. Although it is predicted that apolipoprotein A-I (apoA-I) directly binds to ABCA1, the physiological importance of this interaction is still controversial and the conformation required for apoA-I binding is unclear. In this study, the role of the two nucleotide-binding domains (NBD) of ABCA1 in apoA-I binding was determined by inserting a TEV protease recognition sequence in the linker region of ABCA1. Analyses of ATP binding and occlusion to wild-type ABCA1 and various NBD mutants revealed that ATP binds equally to both NBDs and is hydrolyzed at both NBDs. The interaction with apoA-I and the apoA-I-dependent cholesterol efflux required not only ATP binding but also hydrolysis in both NBDs. NBD mutations and cellular ATP depletion decreased the accessibility of antibodies to a hemagglutinin (HA) epitope that was inserted at position 443 in the extracellular domain (ECD), suggesting that the conformation of ECDs is altered by ATP hydrolysis at both NBDs. These results suggest that ATP hydrolysis at both NBDs induces conformational changes in the ECDs, which are associated with apoA-I binding and cholesterol efflux.http://www.sciencedirect.com/science/article/pii/S0022227520408065ATP binding cassette protein A1apolipoproteinscholesterol effluxHDLtransport |
| spellingShingle | Kohjiro Nagao Kei Takahashi Yuya Azuma Mie Takada Yasuhisa Kimura Michinori Matsuo Noriyuki Kioka Kazumitsu Ueda ATP hydrolysis-dependent conformational changes in the extracellular domain of ABCA1 are associated with apoA-I binding[S] Journal of Lipid Research ATP binding cassette protein A1 apolipoproteins cholesterol efflux HDL transport |
| title | ATP hydrolysis-dependent conformational changes in the extracellular domain of ABCA1 are associated with apoA-I binding[S] |
| title_full | ATP hydrolysis-dependent conformational changes in the extracellular domain of ABCA1 are associated with apoA-I binding[S] |
| title_fullStr | ATP hydrolysis-dependent conformational changes in the extracellular domain of ABCA1 are associated with apoA-I binding[S] |
| title_full_unstemmed | ATP hydrolysis-dependent conformational changes in the extracellular domain of ABCA1 are associated with apoA-I binding[S] |
| title_short | ATP hydrolysis-dependent conformational changes in the extracellular domain of ABCA1 are associated with apoA-I binding[S] |
| title_sort | atp hydrolysis dependent conformational changes in the extracellular domain of abca1 are associated with apoa i binding s |
| topic | ATP binding cassette protein A1 apolipoproteins cholesterol efflux HDL transport |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520408065 |
| work_keys_str_mv | AT kohjironagao atphydrolysisdependentconformationalchangesintheextracellulardomainofabca1areassociatedwithapoaibindings AT keitakahashi atphydrolysisdependentconformationalchangesintheextracellulardomainofabca1areassociatedwithapoaibindings AT yuyaazuma atphydrolysisdependentconformationalchangesintheextracellulardomainofabca1areassociatedwithapoaibindings AT mietakada atphydrolysisdependentconformationalchangesintheextracellulardomainofabca1areassociatedwithapoaibindings AT yasuhisakimura atphydrolysisdependentconformationalchangesintheextracellulardomainofabca1areassociatedwithapoaibindings AT michinorimatsuo atphydrolysisdependentconformationalchangesintheextracellulardomainofabca1areassociatedwithapoaibindings AT noriyukikioka atphydrolysisdependentconformationalchangesintheextracellulardomainofabca1areassociatedwithapoaibindings AT kazumitsuueda atphydrolysisdependentconformationalchangesintheextracellulardomainofabca1areassociatedwithapoaibindings |