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Title: Crystal Structures of SgcE6 and SgcC, the Two-Component Monooxygenase That Catalyzes Hydroxylation of a Carrier ProteinTethered Substrate during the Biosynthesis of the Enediyne Antitumor Antibiotic C-1027 in Streptomyces globisporus

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-beta-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-beta-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 A 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 FADH 2-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, FADH 2-dependent monooxygenase (SgcE6 and SgcC) that catalyzes the hydroxylation of a PCP-tethered substrate.
Authors:
 [1] ;  [1] ;  [2] ;  [3] ;  [1] ;  [1] ;  [2] ;  [4] ;  [5] ;  [3] ;  [3] ;  [1] ;  [3] ;  [2] ;  [1]
  1. The Scripps Research Institute, Jupiter, FL (United States)
  2. Rice Univ., Houston, TX (United States)
  3. Argonne National Lab. (ANL), Argonne, IL (United States)
  4. Univ. of Wisconsin - Madison, Madison, WI (United States)
  5. Rice Univ., Houston, TX (United States); Jaypee Univ. of Information Technology, Pradesh (India)
Publication Date:
Grant/Contract Number:
AC02-06CH11357
Type:
Published Article
Journal Name:
Biochemistry
Additional Journal Information:
Journal Volume: 55; Journal Issue: 36; Journal ID: ISSN 0006-2960
Publisher:
American Chemical Society (ACS)
Research Org:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org:
National Institutes of Health (NIH); USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); Scripps Research Institute
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES
OSTI Identifier:
1324326
Alternate Identifier(s):
OSTI ID: 1416000

Chang, Chin -Yuan, Lohman, Jeremy, 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 ProteinTethered Substrate during the Biosynthesis of the Enediyne Antitumor Antibiotic C-1027 in Streptomyces globisporus. United States: N. p., Web. doi:10.1021/acs.biochem.6b00713.
Chang, Chin -Yuan, Lohman, Jeremy, 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 ProteinTethered Substrate during the Biosynthesis of the Enediyne Antitumor Antibiotic C-1027 in Streptomyces globisporus. United States. doi:10.1021/acs.biochem.6b00713.
Chang, Chin -Yuan, Lohman, Jeremy, 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. 2016. "Crystal Structures of SgcE6 and SgcC, the Two-Component Monooxygenase That Catalyzes Hydroxylation of a Carrier ProteinTethered Substrate during the Biosynthesis of the Enediyne Antitumor Antibiotic C-1027 in Streptomyces globisporus". United States. doi: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 ProteinTethered Substrate during the Biosynthesis of the Enediyne Antitumor Antibiotic C-1027 in Streptomyces globisporus},
author = {Chang, Chin -Yuan and Lohman, Jeremy 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-beta-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-beta-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 A 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 = {2016},
month = {8}
}