TY - JOUR
T1 - Mechanism of intersubunit ketosynthase-dehydratase interaction in polyketide synthases
AU - Jenner, Matthew
AU - Kosol, Simone
AU - Griffiths, Daniel
AU - Prasongpholchai, Panward
AU - Manzi, Lucio
AU - Barrow, Andrew
AU - Moses, John
AU - Oldham, Neil
AU - Lewandowski, Józef
AU - Challis, Gregory L
PY - 2018/3/1
Y1 - 2018/3/1
N2 - Modular polyketide synthases (PKSs) produce numerous structurally complex natural products that have diverse applications in medicine and agriculture. PKSs typically consist of several multienzyme subunits that utilize structurally defined docking domains (DDs) at their N and C termini to ensure correct assembly into functional multiprotein complexes. Here we report a fundamentally different mechanism for subunit assembly in trans-acyltransferase (trans-AT) modular PKSs at the junction between ketosynthase (KS) and dehydratase (DH) domains. This mechanism involves direct interaction of a largely unstructured docking domain (DD) at the C terminus of the KS with the surface of the downstream DH. Acyl transfer assays and mechanism-based crosslinking established that the DD is required for the KS to communicate with the acyl carrier protein appended to the DH. Two distinct regions for binding of the DD to the DH were identified using NMR spectroscopy, carbene footprinting, and mutagenesis, providing a foundation for future elucidation of the molecular basis for interaction specificity.
AB - Modular polyketide synthases (PKSs) produce numerous structurally complex natural products that have diverse applications in medicine and agriculture. PKSs typically consist of several multienzyme subunits that utilize structurally defined docking domains (DDs) at their N and C termini to ensure correct assembly into functional multiprotein complexes. Here we report a fundamentally different mechanism for subunit assembly in trans-acyltransferase (trans-AT) modular PKSs at the junction between ketosynthase (KS) and dehydratase (DH) domains. This mechanism involves direct interaction of a largely unstructured docking domain (DD) at the C terminus of the KS with the surface of the downstream DH. Acyl transfer assays and mechanism-based crosslinking established that the DD is required for the KS to communicate with the acyl carrier protein appended to the DH. Two distinct regions for binding of the DD to the DH were identified using NMR spectroscopy, carbene footprinting, and mutagenesis, providing a foundation for future elucidation of the molecular basis for interaction specificity.
UR - http://www.scopus.com/inward/record.url?scp=85042035197&partnerID=8YFLogxK
U2 - 10.1038/nchembio.2549
DO - 10.1038/nchembio.2549
M3 - Article
AN - SCOPUS:85042035197
SN - 1552-4469
VL - 14
SP - 270
EP - 275
JO - Nature Chemical Biology
JF - Nature Chemical Biology
IS - 3
ER -