Crystal Structures of SgcE6 and SgcC, the Two-Component Monooxygenase That Catalyzes Hydroxylation of a Carrier Protein-Tethered Substrate during the Biosynthesis of the Enediyne Antitumor Antibiotic C-1027 in Streptomyces globisporus
Abstract
C-1027 is a chromoprotein enediyne antitumor antibiotic produced by Streptomyces globisporus. In the last step of biosynthesis of the (S)-3-chloro-5-hydroxy-β-tyrosine moiety of the C-1027 enediyne chromophore, SgcE6 and SgcC compose a two-component monooxygenase that hydroxylates the C-5 position of (S)-3-chloro-β-tyrosine. This two-component monooxygenase is remarkable for two reasons. (i) SgcE6 specifically reacts with FAD and NADH, and (ii) SgcC is active with only the peptidyl carrier protein (PCP)-tethered substrate. To address the molecular details of substrate specificity, we determined the crystal structures of SgcE6 and SgcC at 1.66 and 2.63 Å resolution, respectively. SgcE6 shares a similar β-barrel fold with the class I HpaC-like flavin reductases. A flexible loop near the active site of SgcE6 plays a role in FAD binding, likely by providing sufficient space to accommodate the AMP moiety of FAD, when compared to that of FMN-utilizing homologues. SgcC shows structural similarity to a few other known FADH2-dependent monooxygenases and sheds light on some biochemically but not structurally characterized homologues. In conclusion, the crystal structures reported here provide insights into substrate specificity, and comparison with homologues provides a catalytic mechanism of the two-component, FADH2-dependent monooxygenase (SgcE6 and SgcC) that catalyzes the hydroxylation of a PCP-tethered substrate.
- Authors:
-
- Department of Chemistry, The Scripps Research Institute, Jupiter, Florida 33458, United States
- BioScience at Rice and Department of Chemistry, Rice University, Houston, Texas 77251, United States
- Midwest Center for Structural Genomics and Structural Biology Center, Biosciences Division, Argonne National Laboratory, Argonne, Illinois 60439, United States
- Department of Biochemistry, University of Wisconsin—Madison, Madison, Wisconsin 53705, United States
- BioScience at Rice and Department of Chemistry, Rice University, Houston, Texas 77251, United States, Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Himachal Pradesh, India 173234
- Publication Date:
- Research Org.:
- Scripps Research Inst., Jupiter, FL (United States); Argonne National Laboratory (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH); Scripps Research Institute
- Contributing Org.:
- Structural Biology Center beamlines at the Advanced Photon Source
- OSTI Identifier:
- 1324326
- Alternate Identifier(s):
- OSTI ID: 1326642; OSTI ID: 1416000
- Grant/Contract Number:
- AC02-06CH11357; GM098248; GM109456; GM094585; CA078747
- Resource Type:
- Published Article
- Journal Name:
- Biochemistry
- Additional Journal Information:
- Journal Name: Biochemistry Journal Volume: 55 Journal Issue: 36; Journal ID: ISSN 0006-2960
- Publisher:
- American Chemical Society
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES
Citation Formats
Chang, Chin-Yuan, Lohman, Jeremy R., Cao, Hongnan, Tan, Kemin, Rudolf, Jeffrey D., Ma, Ming, Xu, Weijun, Bingman, Craig A., Yennamalli, Ragothaman M., Bigelow, Lance, Babnigg, Gyorgy, Yan, Xiaohui, Joachimiak, Andrzej, Phillips, Jr., George N., and Shen, Ben. Crystal Structures of SgcE6 and SgcC, the Two-Component Monooxygenase That Catalyzes Hydroxylation of a Carrier Protein-Tethered Substrate during the Biosynthesis of the Enediyne Antitumor Antibiotic C-1027 in Streptomyces globisporus. United States: N. p., 2016.
Web. doi:10.1021/acs.biochem.6b00713.
Chang, Chin-Yuan, Lohman, Jeremy R., Cao, Hongnan, Tan, Kemin, Rudolf, Jeffrey D., Ma, Ming, Xu, Weijun, Bingman, Craig A., Yennamalli, Ragothaman M., Bigelow, Lance, Babnigg, Gyorgy, Yan, Xiaohui, Joachimiak, Andrzej, Phillips, Jr., George N., & Shen, Ben. Crystal Structures of SgcE6 and SgcC, the Two-Component Monooxygenase That Catalyzes Hydroxylation of a Carrier Protein-Tethered Substrate during the Biosynthesis of the Enediyne Antitumor Antibiotic C-1027 in Streptomyces globisporus. United States. https://doi.org/10.1021/acs.biochem.6b00713
Chang, Chin-Yuan, Lohman, Jeremy R., Cao, Hongnan, Tan, Kemin, Rudolf, Jeffrey D., Ma, Ming, Xu, Weijun, Bingman, Craig A., Yennamalli, Ragothaman M., Bigelow, Lance, Babnigg, Gyorgy, Yan, Xiaohui, Joachimiak, Andrzej, Phillips, Jr., George N., and Shen, Ben. Thu .
"Crystal Structures of SgcE6 and SgcC, the Two-Component Monooxygenase That Catalyzes Hydroxylation of a Carrier Protein-Tethered Substrate during the Biosynthesis of the Enediyne Antitumor Antibiotic C-1027 in Streptomyces globisporus". United States. https://doi.org/10.1021/acs.biochem.6b00713.
@article{osti_1324326,
title = {Crystal Structures of SgcE6 and SgcC, the Two-Component Monooxygenase That Catalyzes Hydroxylation of a Carrier Protein-Tethered Substrate during the Biosynthesis of the Enediyne Antitumor Antibiotic C-1027 in Streptomyces globisporus},
author = {Chang, Chin-Yuan and Lohman, Jeremy R. and Cao, Hongnan and Tan, Kemin and Rudolf, Jeffrey D. and Ma, Ming and Xu, Weijun and Bingman, Craig A. and Yennamalli, Ragothaman M. and Bigelow, Lance and Babnigg, Gyorgy and Yan, Xiaohui and Joachimiak, Andrzej and Phillips, Jr., George N. and Shen, Ben},
abstractNote = {C-1027 is a chromoprotein enediyne antitumor antibiotic produced by Streptomyces globisporus. In the last step of biosynthesis of the (S)-3-chloro-5-hydroxy-β-tyrosine moiety of the C-1027 enediyne chromophore, SgcE6 and SgcC compose a two-component monooxygenase that hydroxylates the C-5 position of (S)-3-chloro-β-tyrosine. This two-component monooxygenase is remarkable for two reasons. (i) SgcE6 specifically reacts with FAD and NADH, and (ii) SgcC is active with only the peptidyl carrier protein (PCP)-tethered substrate. To address the molecular details of substrate specificity, we determined the crystal structures of SgcE6 and SgcC at 1.66 and 2.63 Å resolution, respectively. SgcE6 shares a similar β-barrel fold with the class I HpaC-like flavin reductases. A flexible loop near the active site of SgcE6 plays a role in FAD binding, likely by providing sufficient space to accommodate the AMP moiety of FAD, when compared to that of FMN-utilizing homologues. SgcC shows structural similarity to a few other known FADH2-dependent monooxygenases and sheds light on some biochemically but not structurally characterized homologues. In conclusion, the crystal structures reported here provide insights into substrate specificity, and comparison with homologues provides a catalytic mechanism of the two-component, FADH2-dependent monooxygenase (SgcE6 and SgcC) that catalyzes the hydroxylation of a PCP-tethered substrate.},
doi = {10.1021/acs.biochem.6b00713},
journal = {Biochemistry},
number = 36,
volume = 55,
place = {United States},
year = {Thu Sep 01 00:00:00 EDT 2016},
month = {Thu Sep 01 00:00:00 EDT 2016}
}
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https://doi.org/10.1021/acs.biochem.6b00713
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