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Title: Structure–Function Analysis of the Extended Conformation of a Polyketide Synthase Module

Abstract

Catalytic modules of assembly-line polyketide synthases (PKSs) have previously been observed in two very different conformations—an “extended” architecture and an “arch-shaped” architecture—although the catalytic relevance of neither has been directly established. By the use of a fully human naïve antigen-binding fragment (Fab) library, a high-affinity antibody was identified that bound to the extended conformation of a PKS module, as verified by X-ray crystallography and tandem size-exclusion chromatography–small-angle X-ray scattering (SEC–SAXS). Kinetic analysis proved that this antibody-stabilized module conformation was fully competent for catalysis of intermodular polyketide chain translocation as well as intramodular polyketide chain elongation and functional group modification of a growing polyketide chain. Furthermore, the extended conformation of a PKS module is fully competent for all of its essential catalytic functions.

Authors:
 [1];  [2];  [2];  [1]; ORCiD logo [1];  [3]; ORCiD logo [4];  [4]; ORCiD logo [5]; ORCiD logo [2]; ORCiD logo [1]
  1. Stanford Univ., Stanford, CA (United States)
  2. University of California San Francisco, San Francisco, CA (United States)
  3. Stanford Univ., Stanford, CA (United States); SLAC National Accelerator Lab., Stanford Univ., Menlo Park, CA (United States)
  4. SLAC National Accelerator Lab., Stanford Univ., Menlo Park, CA (United States)
  5. Brown Univ., Providence, RI (United States)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1476142
Grant/Contract Number:  
AC02-76SF00515; P41CA196276; GM022172; GM104659; GM087934
Resource Type:
Accepted Manuscript
Journal Name:
Journal of the American Chemical Society
Additional Journal Information:
Journal Volume: 140; Journal Issue: 21; Journal ID: ISSN 0002-7863
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY

Citation Formats

Li, Xiuyuan, Sevillano, Natalia, La Greca, Florencia, Deis, Lindsay, Liu, Yu -Chen, Deller, Marc C., Mathews, Irimpan I., Matsui, Tsutomu, Cane, David E., Craik, Charles S., and Khosla, Chaitan. Structure–Function Analysis of the Extended Conformation of a Polyketide Synthase Module. United States: N. p., 2018. Web. doi:10.1021/jacs.8b02100.
Li, Xiuyuan, Sevillano, Natalia, La Greca, Florencia, Deis, Lindsay, Liu, Yu -Chen, Deller, Marc C., Mathews, Irimpan I., Matsui, Tsutomu, Cane, David E., Craik, Charles S., & Khosla, Chaitan. Structure–Function Analysis of the Extended Conformation of a Polyketide Synthase Module. United States. https://doi.org/10.1021/jacs.8b02100
Li, Xiuyuan, Sevillano, Natalia, La Greca, Florencia, Deis, Lindsay, Liu, Yu -Chen, Deller, Marc C., Mathews, Irimpan I., Matsui, Tsutomu, Cane, David E., Craik, Charles S., and Khosla, Chaitan. Tue . "Structure–Function Analysis of the Extended Conformation of a Polyketide Synthase Module". United States. https://doi.org/10.1021/jacs.8b02100. https://www.osti.gov/servlets/purl/1476142.
@article{osti_1476142,
title = {Structure–Function Analysis of the Extended Conformation of a Polyketide Synthase Module},
author = {Li, Xiuyuan and Sevillano, Natalia and La Greca, Florencia and Deis, Lindsay and Liu, Yu -Chen and Deller, Marc C. and Mathews, Irimpan I. and Matsui, Tsutomu and Cane, David E. and Craik, Charles S. and Khosla, Chaitan},
abstractNote = {Catalytic modules of assembly-line polyketide synthases (PKSs) have previously been observed in two very different conformations—an “extended” architecture and an “arch-shaped” architecture—although the catalytic relevance of neither has been directly established. By the use of a fully human naïve antigen-binding fragment (Fab) library, a high-affinity antibody was identified that bound to the extended conformation of a PKS module, as verified by X-ray crystallography and tandem size-exclusion chromatography–small-angle X-ray scattering (SEC–SAXS). Kinetic analysis proved that this antibody-stabilized module conformation was fully competent for catalysis of intermodular polyketide chain translocation as well as intramodular polyketide chain elongation and functional group modification of a growing polyketide chain. Furthermore, the extended conformation of a PKS module is fully competent for all of its essential catalytic functions.},
doi = {10.1021/jacs.8b02100},
journal = {Journal of the American Chemical Society},
number = 21,
volume = 140,
place = {United States},
year = {2018},
month = {5}
}

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Figures / Tables:

Figure 1 Figure 1: (A) Bimodular and (B) trirnodular derivatives of DEBS. Domains are shown as spheres. KS = ketosynthase; AT = acyl transferase; ACP = acyl carrier protein; L = KS-AT linker domain; KR = ketoreductase; KR0 = redox-inactive, epimerase-active KR homologue; TE = thioesterase. Blue tabs depict docking domains thatmore » facilitate non-covalent association of successive modules. Module 2 domains are colored for ease of comparison with Figure 2.« less

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