Nonribosomal peptide synthetases (NRPSs) use repeats of domains to tether substrates onto carrier protein domains (CP) and assemble them into complex products through intervening condensation or cyclization domains. A dynamic domain architecture has hampered NRPS engineering because it is unclear whether and how catalytic steps required for synthesis drive transient domain interactions. Here, we demonstrate experimentally that large scale structural dynamics within a 52 kDa cyclization domain sense the attachment of substrates to its partner CP and promote binding while opening an allosteric path to a remote binding site. The demonstration relies on a novel nuclear magnetic resonance experimental design, in which we monitor the molecular response of a protein towards covalent modifications of its binding partner <i>in situ</i>. Our results exemplify how structural dynamics within proteins can couple substrate recognition with active site remodeling, and how they can promote both local and remote protein communication.

Structural dynamics mediate tandem substrate recognition and allostery in a peptide synthetase domain

ACS Fall 2021

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