dc.contributor.author | Saenz-Vash, Veronica | |
dc.contributor.author | Romer, Katherine A. | |
dc.contributor.author | Xu, Shangzhe | |
dc.contributor.author | Yesilaltay, Ayce | |
dc.contributor.author | Krieger, Monty | |
dc.contributor.author | Yu, Miao | |
dc.contributor.author | Nieland, Thomas J | |
dc.contributor.author | Penman, Marsha L | |
dc.contributor.author | Carr, Steven A | |
dc.date.accessioned | 2012-02-01T21:24:15Z | |
dc.date.available | 2012-02-01T21:24:15Z | |
dc.date.issued | 2011-07 | |
dc.date.submitted | 2011-04 | |
dc.identifier.issn | 0027-8424 | |
dc.identifier.issn | 1091-6490 | |
dc.identifier.uri | http://hdl.handle.net/1721.1/69003 | |
dc.description.abstract | The HDL receptor, scavenger receptor, class B, type I (SR-BI), is a homooligomeric cell surface glycoprotein that controls HDL structure and metabolism by mediating the cellular selective uptake of lipids, mainly cholesteryl esters, from HDL. The mechanism underlying SR-BI-mediated lipid transfer, which differs from classic receptor-mediated endocytosis, involves a two-step process (binding followed by lipid transport) that is poorly understood. Our previous structure/activity analysis of the small-molecule inhibitor blocker of lipid transport 1 (BLT-1), which potently (IC[subscript 50] ∼ 50 nM) blocks SR-BI-mediated lipid transport, established that the sulfur in BLT-1’s thiosemicarbazone moiety was essential for activity. Here we show that BLT-1 is an irreversible inhibitor of SR-BI, raising the possibility that cysteine(s) in SR-BI interact with BLT-1. Mass spectrometric analysis of purified SR-BI showed two of its six exoplasmic cysteines have free thiol groups (Cys251 and Cys384). Converting Cys384 (but not Cys251) to serine resulted in complete BLT-1 insensitivity, establishing that the unique molecular target of BLT-1 inhibition of cellular SR-BI dependent lipid transport is SR-BI itself. The C384S substitution reduced the receptor’s intrinsic lipid uptake activity by approximately 60% without dramatically altering its surface expression, homooligomerization, or HDL binding. Thus, a small-molecule screening approach identified a key residue in SR-BI involved in lipid transport, providing a powerful springboard into the analyses of the structure and mechanism of SR-BI, and highlighting the power of this approach for such analyses. | en_US |
dc.description.sponsorship | Broad Institute of MIT and Harvard. Proteomics Platform | en_US |
dc.description.sponsorship | National Cancer Institute (U.S.) | en_US |
dc.language.iso | en_US | |
dc.publisher | Proceedings of the National Academy of Sciences (PNAS) | en_US |
dc.relation.isversionof | http://dx.doi.org/10.1073/pnas.1109078108 | en_US |
dc.rights | Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. | en_US |
dc.source | PNAS | en_US |
dc.title | Exoplasmic cysteine Cys384 of the HDL receptor SR-BI is critical for its sensitivity to a small-molecule inhibitor and normal lipid transport activity | en_US |
dc.type | Article | en_US |
dc.identifier.citation | Yu, M. et al. “Exoplasmic cysteine Cys384 of the HDL receptor SR-BI is critical for its sensitivity to a small-molecule inhibitor and normal lipid transport activity.” Proceedings of the National Academy of Sciences 108.30 (2011): 12243-12248. Web. 1 Feb. 2012. | en_US |
dc.contributor.department | Massachusetts Institute of Technology. Computational and Systems Biology Program | en_US |
dc.contributor.department | Massachusetts Institute of Technology. Department of Biology | en_US |
dc.contributor.department | Koch Institute for Integrative Cancer Research at MIT | en_US |
dc.contributor.approver | Krieger, Monty | |
dc.contributor.mitauthor | Yua, Miao | |
dc.contributor.mitauthor | Romer, Katherine A. | |
dc.contributor.mitauthor | Nieland, Thomas J. | |
dc.contributor.mitauthor | Xu, Shangzhe | |
dc.contributor.mitauthor | Penman, Marsha L. | |
dc.contributor.mitauthor | Yesilaltay, Ayce | |
dc.contributor.mitauthor | Carr, Steven A. | |
dc.contributor.mitauthor | Krieger, Monty | |
dc.relation.journal | Proceedings of the National Academy of Sciences | en_US |
dc.eprint.version | Final published version | en_US |
dc.type.uri | http://purl.org/eprint/type/JournalArticle | en_US |
eprint.status | http://purl.org/eprint/status/PeerReviewed | en_US |
dspace.orderedauthors | Yu, M.; Romer, K. A.; Nieland, T. J. F.; Xu, S.; Saenz-Vash, V.; Penman, M.; Yesilaltay, A.; Carr, S. A.; Krieger, M. | en |
dc.identifier.orcid | https://orcid.org/0000-0003-4541-5181 | |
dc.identifier.orcid | https://orcid.org/0000-0003-2673-1672 | |
dc.identifier.orcid | https://orcid.org/0000-0002-7203-4299 | |
dc.identifier.orcid | https://orcid.org/0000-0002-5503-182X | |
dc.identifier.orcid | https://orcid.org/0000-0001-9905-5316 | |
mit.license | PUBLISHER_POLICY | en_US |
mit.metadata.status | Complete | |