Structure of human Fe–S assembly subcomplex reveals unexpected cysteine desulfurase architecture and acyl-ACP–ISD11 interactions
In eukaryotes, sulfur is mobilized for incorporation into multiple biosynthetic pathways by a cysteine desulfurase complex that consists of a catalytic subunit (NFS1), LYR protein (ISD11), and acyl carrier protein (ACP). This NFS1-ISD11-ACP (SDA) complex forms the core of the iron-sulfur (Fe-S) asse...
| Main Authors: | , , , , , , , |
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| Format: | Article |
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National Academy of Sciences (U.S.)
2018
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| Online Access: | http://hdl.handle.net/1721.1/114922 https://orcid.org/0000-0002-4285-6128 https://orcid.org/0000-0001-5486-2755 |
| _version_ | 1811094770245697536 |
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| author | Cory, Seth A. Van Vranken, Jonathan G. Brignole, Edward J. Patra, Shachin Winge, Dennis R. Drennan, Catherine L. Rutter, Jared Barondeau, David P. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Cory, Seth A. Van Vranken, Jonathan G. Brignole, Edward J. Patra, Shachin Winge, Dennis R. Drennan, Catherine L. Rutter, Jared Barondeau, David P. |
| author_sort | Cory, Seth A. |
| collection | MIT |
| description | In eukaryotes, sulfur is mobilized for incorporation into multiple biosynthetic pathways by a cysteine desulfurase complex that consists of a catalytic subunit (NFS1), LYR protein (ISD11), and acyl carrier protein (ACP). This NFS1-ISD11-ACP (SDA) complex forms the core of the iron-sulfur (Fe-S) assembly complex and associates with assembly proteins ISCU2, frataxin (FXN), and ferredoxin to synthesize Fe-S clusters. Here we present crystallographic and electron microscopic structures of the SDA complex coupled to enzyme kinetic and cell-based studies to provide structure-function properties of a mitochondrial cysteine desulfurase. Unlike prokaryotic cysteine desulfurases, the SDA structure adopts an unexpected architecture in which a pair of ISD11 subunits form the dimeric core of the SDA complex, which clarifies the critical role of ISD11 in eukaryotic assemblies. The different quaternary structure results in an incompletely formed substrate channel and solvent-exposed pyridoxal 5'-phosphate cofactor and provides a rationale for the allosteric activator function of FXN in eukaryotic systems. The structure also reveals the 4'-phosphopantetheine-conjugated acyl-group of ACP occupies the hydrophobic core of ISD11, explaining the basis of ACP stabilization. The unexpected architecture for the SDA complex provides a framework for understanding interactions with acceptor proteins for sulfur-containing biosynthetic pathways, elucidating mechanistic details of eukaryotic F e-S cluster biosynthesis, and clarifying how defects in Fe-S cluster assembly lead to diseases such as Friedreich's ataxia. Moreover, our results support a lock-and-key model in which LYR proteins associate with acyl-ACP as a mechanism for fatty acid biosynthesis to coordinate the expression, Fe-S cofactor maturation, and activity of the respiratory complexes. Keywords: LYR; ACP; iron-sulfur cluster; PLP; frataxin |
| first_indexed | 2024-09-23T16:04:28Z |
| format | Article |
| id | mit-1721.1/114922 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T16:04:28Z |
| publishDate | 2018 |
| publisher | National Academy of Sciences (U.S.) |
| record_format | dspace |
| spelling | mit-1721.1/1149222022-09-29T18:05:41Z Structure of human Fe–S assembly subcomplex reveals unexpected cysteine desulfurase architecture and acyl-ACP–ISD11 interactions Cory, Seth A. Van Vranken, Jonathan G. Brignole, Edward J. Patra, Shachin Winge, Dennis R. Drennan, Catherine L. Rutter, Jared Barondeau, David P. Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemistry Brignole, Edward J. Drennan, Catherine L. In eukaryotes, sulfur is mobilized for incorporation into multiple biosynthetic pathways by a cysteine desulfurase complex that consists of a catalytic subunit (NFS1), LYR protein (ISD11), and acyl carrier protein (ACP). This NFS1-ISD11-ACP (SDA) complex forms the core of the iron-sulfur (Fe-S) assembly complex and associates with assembly proteins ISCU2, frataxin (FXN), and ferredoxin to synthesize Fe-S clusters. Here we present crystallographic and electron microscopic structures of the SDA complex coupled to enzyme kinetic and cell-based studies to provide structure-function properties of a mitochondrial cysteine desulfurase. Unlike prokaryotic cysteine desulfurases, the SDA structure adopts an unexpected architecture in which a pair of ISD11 subunits form the dimeric core of the SDA complex, which clarifies the critical role of ISD11 in eukaryotic assemblies. The different quaternary structure results in an incompletely formed substrate channel and solvent-exposed pyridoxal 5'-phosphate cofactor and provides a rationale for the allosteric activator function of FXN in eukaryotic systems. The structure also reveals the 4'-phosphopantetheine-conjugated acyl-group of ACP occupies the hydrophobic core of ISD11, explaining the basis of ACP stabilization. The unexpected architecture for the SDA complex provides a framework for understanding interactions with acceptor proteins for sulfur-containing biosynthetic pathways, elucidating mechanistic details of eukaryotic F e-S cluster biosynthesis, and clarifying how defects in Fe-S cluster assembly lead to diseases such as Friedreich's ataxia. Moreover, our results support a lock-and-key model in which LYR proteins associate with acyl-ACP as a mechanism for fatty acid biosynthesis to coordinate the expression, Fe-S cofactor maturation, and activity of the respiratory complexes. Keywords: LYR; ACP; iron-sulfur cluster; PLP; frataxin 2018-04-24T14:07:42Z 2018-04-24T14:07:42Z 2017-06 2017-02 2018-04-20T16:02:58Z Article http://purl.org/eprint/type/ConferencePaper 0027-8424 1091-6490 http://hdl.handle.net/1721.1/114922 Cory, Seth A. et al. “Structure of Human Fe–S Assembly Subcomplex Reveals Unexpected Cysteine Desulfurase Architecture and Acyl-ACP–ISD11 Interactions.” Proceedings of the National Academy of Sciences 114, 27 (June 2017): E5325–E5334 © 2017 The Authors https://orcid.org/0000-0002-4285-6128 https://orcid.org/0000-0001-5486-2755 http://dx.doi.org/10.1073/PNAS.1702849114 Proceedings of the National Academy of Sciences 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. application/pdf National Academy of Sciences (U.S.) National Academy of Sciences |
| spellingShingle | Cory, Seth A. Van Vranken, Jonathan G. Brignole, Edward J. Patra, Shachin Winge, Dennis R. Drennan, Catherine L. Rutter, Jared Barondeau, David P. Structure of human Fe–S assembly subcomplex reveals unexpected cysteine desulfurase architecture and acyl-ACP–ISD11 interactions |
| title | Structure of human Fe–S assembly subcomplex reveals unexpected cysteine desulfurase architecture and acyl-ACP–ISD11 interactions |
| title_full | Structure of human Fe–S assembly subcomplex reveals unexpected cysteine desulfurase architecture and acyl-ACP–ISD11 interactions |
| title_fullStr | Structure of human Fe–S assembly subcomplex reveals unexpected cysteine desulfurase architecture and acyl-ACP–ISD11 interactions |
| title_full_unstemmed | Structure of human Fe–S assembly subcomplex reveals unexpected cysteine desulfurase architecture and acyl-ACP–ISD11 interactions |
| title_short | Structure of human Fe–S assembly subcomplex reveals unexpected cysteine desulfurase architecture and acyl-ACP–ISD11 interactions |
| title_sort | structure of human fe s assembly subcomplex reveals unexpected cysteine desulfurase architecture and acyl acp isd11 interactions |
| url | http://hdl.handle.net/1721.1/114922 https://orcid.org/0000-0002-4285-6128 https://orcid.org/0000-0001-5486-2755 |
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