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Solution structure and proposed domain–domain recognition interface of an acyl carrier protein domain from a modular polyketide synthase
Author(s) -
Alekseyev Viktor Y.,
Liu Corey W.,
Cane David E.,
Puglisi Joseph D.,
Khosla Chaitan
Publication year - 2007
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.073011407
Subject(s) - polyketide , polyketide synthase , acyl carrier protein , chemistry , stereochemistry , crystallography , homology modeling , nuclear magnetic resonance spectroscopy , protein structure , helix (gastropod) , biosynthesis , biology , biochemistry , enzyme , ecology , snail
Polyketides are a medicinally important class of natural products. The architecture of modular polyketide synthases (PKSs), composed of multiple covalently linked domains grouped into modules, provides an attractive framework for engineering novel polyketide‐producing assemblies. However, impaired domain–domain interactions can compromise the efficiency of engineered polyketide biosynthesis. To facilitate the study of these domain–domain interactions, we have used nuclear magnetic resonance (NMR) spectroscopy to determine the first solution structure of an acyl carrier protein (ACP) domain from a modular PKS, 6‐deoxyerythronolide B synthase (DEBS). The tertiary fold of this 10‐kD domain is a three‐helical bundle; an additional short helix in the second loop also contributes to the core helical packing. Superposition of residues 14–94 of the ensemble on the mean structure yields an average atomic RMSD of 0.64 ± 0.09 Å for the backbone atoms (1.21 ± 0.13 Å for all non‐hydrogen atoms). The three major helices superimpose with a backbone RMSD of 0.48 ± 0.10 Å (0.99 ± 0.11 Å for non‐hydrogen atoms). Based on this solution structure, homology models were constructed for five other DEBS ACP domains. Comparison of their steric and electrostatic surfaces at the putative interaction interface (centered on helix II) suggests a model for protein–protein recognition of ACP domains, consistent with the previously observed specificity. Site‐directed mutagenesis experiments indicate that two of the identified residues influence the specificity of ACP recognition.