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TitleStructural basis for intermodular communication in assembly-line polyketide biosynthesis.
Journal, issue, pagesNat Chem Biol, Year 2024
Publish dateAug 23, 2024
AuthorsDillon P Cogan / Alexander M Soohoo / Muyuan Chen / Yan Liu / Krystal L Brodsky / Chaitan Khosla /
PubMed AbstractAssembly-line polyketide synthases (PKSs) are modular multi-enzyme systems with considerable potential for genetic reprogramming. Understanding how they selectively transport biosynthetic ...Assembly-line polyketide synthases (PKSs) are modular multi-enzyme systems with considerable potential for genetic reprogramming. Understanding how they selectively transport biosynthetic intermediates along a defined sequence of active sites could be harnessed to rationally alter PKS product structures. To investigate functional interactions between PKS catalytic and substrate acyl carrier protein (ACP) domains, we employed a bifunctional reagent to crosslink transient domain-domain interfaces of a prototypical assembly line, the 6-deoxyerythronolide B synthase, and resolved their structures by single-particle cryogenic electron microscopy (cryo-EM). Together with statistical per-particle image analysis of cryo-EM data, we uncovered interactions between ketosynthase (KS) and ACP domains that discriminate between intra-modular and inter-modular communication while reinforcing the relevance of conformational asymmetry during the catalytic cycle. Our findings provide a foundation for the structure-based design of hybrid PKSs comprising biosynthetic modules from different naturally occurring assembly lines.
External linksNat Chem Biol / PubMed:39179672
MethodsEM (single particle)
Resolution3.05 - 3.73 Å
Structure data

41305

8tjn

EMDB-41305, PDB-8tjn:
Crosslinked 6-deoxyerythronolide B synthase (DEBS) Module 1 in complex with antibody fragment 1B2: Crosslinked State 1
Method: EM (single particle) / Resolution: 3.73 Å

41306

8tjo

EMDB-41306, PDB-8tjo:
Crosslinked 6-deoxyerythronolide B synthase (DEBS) Module 1 in complex with antibody fragment 1B2: Crosslinked Intra-State 1
Method: EM (single particle) / Resolution: 3.61 Å

41307

8tjp

EMDB-41307, PDB-8tjp:
KS-AT core of 6-deoxyerythronolide B synthase (DEBS) Module 3 crosslinked with its elongation ACP partner
Method: EM (single particle) / Resolution: 3.71 Å

41355

8tko

EMDB-41355, PDB-8tko:
KS-AT core of 6-deoxyerythronolide B synthase (DEBS) Module 3 crosslinked with its translocation ACP partner of Module 2
Method: EM (single particle) / Resolution: 3.05 Å

41495

8tpw

EMDB-41495, PDB-8tpw:
Crosslinked 6-deoxyerythronolide B synthase (DEBS) Module 3 in complex with antibody fragment 1B2: cis-oriented 1B2 and ACP
Method: EM (single particle) / Resolution: 3.46 Å

41496

8tpx

EMDB-41496, PDB-8tpx:
Crosslinked 6-deoxyerythronolide B synthase (DEBS) Module 3 in complex with antibody fragment 1B2: trans-oriented 1B2 and ACP
Method: EM (single particle) / Resolution: 3.4 Å

Source
  • saccharopolyspora erythraea (bacteria)
  • homo sapiens (human)
KeywordsBIOSYNTHETIC PROTEIN/IMMUNE SYSTEM / polyketide synthase / antibody / BIOSYNTHETIC PROTEIN-IMMUNE SYSTEM complex / BIOSYNTHETIC PROTEIN

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