Crystal structure of the acyltransferase domain of the iterative polyketide synthase in enediyne biosynthesis
Biosynthesis of the enediyne natural product dynemicin in Micromonospora chersina is initiated by DynE8, a highly reducing iterative type I polyketide synthase that assembles polyketide intermediates from the acetate units derived solely from malonyl-CoA. To understand the substrate specificity and...
| Main Authors: | , , , , , , |
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| Format: | Journal Article |
| Language: | English |
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2013
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| Online Access: | https://hdl.handle.net/10356/104167 http://hdl.handle.net/10220/16984 |
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| author | Liew, Chong Wai Nilsson, Martina Chen, Ming Wei Sun, Huihua Cornvik, Tobias Carl Liang, Zhao-Xun Lescar, Julien |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Liew, Chong Wai Nilsson, Martina Chen, Ming Wei Sun, Huihua Cornvik, Tobias Carl Liang, Zhao-Xun Lescar, Julien |
| author_sort | Liew, Chong Wai |
| collection | NTU |
| description | Biosynthesis of the enediyne natural product dynemicin in Micromonospora chersina is initiated by DynE8, a highly reducing iterative type I polyketide synthase that assembles polyketide intermediates from the acetate units derived solely from malonyl-CoA. To understand the substrate specificity and the evolutionary relationship between the acyltransferase (AT) domains of DynE8, fatty acid synthase, and modular polyketide synthases, we overexpressed a 44-kDa fragment of DynE8 (hereafter named ATDYN10) encompassing its entire AT domain and the adjacent linker domain. The crystal structure at 1.4 Å resolution unveils a α/β hydrolase and a ferredoxin-like subdomain with the Ser-His catalytic dyad located in the cleft between the two subdomains. The linker domain also adopts a α/β fold abutting the AT catalytic domain. Co-crystallization with malonyl-CoA yielded a malonyl-enzyme covalent complex that most likely represents the acyl-enzyme intermediate. The structure explains the preference for malonyl-CoA with a conserved arginine orienting the carboxylate group of malonate and several nonpolar residues that preclude α-alkyl malonyl-CoA binding. Co-crystallization with acetyl-CoA revealed two noncovalently bound acetates generated by the enzymatic hydrolysis of acetyl-CoA that acts as an inhibitor for DynE8. This suggests that the AT domain can upload the acyl groups from either malonyl-CoA or acetyl-CoA onto the catalytic Ser651 residue. However, although the malonyl group can be transferred to the acyl carrier protein domain, transfer of the acetyl group to the acyl carrier protein domain is suppressed. Local structural differences may account for the different stability of the acyl-enzyme intermediates. |
| first_indexed | 2024-10-01T05:53:27Z |
| format | Journal Article |
| id | ntu-10356/104167 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T05:53:27Z |
| publishDate | 2013 |
| record_format | dspace |
| spelling | ntu-10356/1041672022-02-16T16:28:36Z Crystal structure of the acyltransferase domain of the iterative polyketide synthase in enediyne biosynthesis Liew, Chong Wai Nilsson, Martina Chen, Ming Wei Sun, Huihua Cornvik, Tobias Carl Liang, Zhao-Xun Lescar, Julien School of Biological Sciences DRNTU::Science::Biological sciences Biosynthesis of the enediyne natural product dynemicin in Micromonospora chersina is initiated by DynE8, a highly reducing iterative type I polyketide synthase that assembles polyketide intermediates from the acetate units derived solely from malonyl-CoA. To understand the substrate specificity and the evolutionary relationship between the acyltransferase (AT) domains of DynE8, fatty acid synthase, and modular polyketide synthases, we overexpressed a 44-kDa fragment of DynE8 (hereafter named ATDYN10) encompassing its entire AT domain and the adjacent linker domain. The crystal structure at 1.4 Å resolution unveils a α/β hydrolase and a ferredoxin-like subdomain with the Ser-His catalytic dyad located in the cleft between the two subdomains. The linker domain also adopts a α/β fold abutting the AT catalytic domain. Co-crystallization with malonyl-CoA yielded a malonyl-enzyme covalent complex that most likely represents the acyl-enzyme intermediate. The structure explains the preference for malonyl-CoA with a conserved arginine orienting the carboxylate group of malonate and several nonpolar residues that preclude α-alkyl malonyl-CoA binding. Co-crystallization with acetyl-CoA revealed two noncovalently bound acetates generated by the enzymatic hydrolysis of acetyl-CoA that acts as an inhibitor for DynE8. This suggests that the AT domain can upload the acyl groups from either malonyl-CoA or acetyl-CoA onto the catalytic Ser651 residue. However, although the malonyl group can be transferred to the acyl carrier protein domain, transfer of the acetyl group to the acyl carrier protein domain is suppressed. Local structural differences may account for the different stability of the acyl-enzyme intermediates. 2013-10-28T08:43:53Z 2019-12-06T21:27:39Z 2013-10-28T08:43:53Z 2019-12-06T21:27:39Z 2012 2012 Journal Article Liew, C. W., Nilsson, M., Chen, M. W., Sun, H., Cornvik, T. C., Liang, Z.-X. et al. (2012). Crystal structure of the acyltransferase domain of the iterative polyketide synthase in enediyne biosynthesis. The journal of biological chemistry, 287, 23203-23215. https://hdl.handle.net/10356/104167 http://hdl.handle.net/10220/16984 10.1074/jbc.M112.362210 22589546 en The journal of biological chemistry |
| spellingShingle | DRNTU::Science::Biological sciences Liew, Chong Wai Nilsson, Martina Chen, Ming Wei Sun, Huihua Cornvik, Tobias Carl Liang, Zhao-Xun Lescar, Julien Crystal structure of the acyltransferase domain of the iterative polyketide synthase in enediyne biosynthesis |
| title | Crystal structure of the acyltransferase domain of the iterative polyketide synthase in enediyne biosynthesis |
| title_full | Crystal structure of the acyltransferase domain of the iterative polyketide synthase in enediyne biosynthesis |
| title_fullStr | Crystal structure of the acyltransferase domain of the iterative polyketide synthase in enediyne biosynthesis |
| title_full_unstemmed | Crystal structure of the acyltransferase domain of the iterative polyketide synthase in enediyne biosynthesis |
| title_short | Crystal structure of the acyltransferase domain of the iterative polyketide synthase in enediyne biosynthesis |
| title_sort | crystal structure of the acyltransferase domain of the iterative polyketide synthase in enediyne biosynthesis |
| topic | DRNTU::Science::Biological sciences |
| url | https://hdl.handle.net/10356/104167 http://hdl.handle.net/10220/16984 |
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