6M01

The structure of HitB-HitD complex

Summary for 6M01

• Entry DOI: 10.2210/pdb6m01/pdb
• Descriptor: Putative ATP-dependent b-aminoacyl-ACP synthetase, Putative ACP, ADENOSINE-5'-DIPHOSPHATE, ... (6 entities in total)
• Functional Keywords: hitachimycin, polyketide biosynthesis, carrier protein, adenylation, atp binding, ligase
• Biological source: Embleya scabrispora; More
• Total number of polymer chains: 2
• Total formula weight: 71646.81

Authors

Miyanaga, A.,Kurihara, S.,Kudo, F.,Eguchi, T. (deposition date: 2020-02-19, release date: 2020-07-08, Last modification date: 2024-10-23)

Primary citation

Miyanaga, A.,Kurihara, S.,Chisuga, T.,Kudo, F.,Eguchi, T.
Structural Characterization of Complex of Adenylation Domain and Carrier Protein by Using Pantetheine Cross-Linking Probe.
Acs Chem.Biol., 15:1808-1812, 2020

PubMed Abstract: Adenylation domains (A-domains) are responsible for selective incorporation of carboxylic acid substrates in the biosynthesis of various natural products. Each A-domain must recognize a cognate carrier protein (CP) for functional substrate transfer. The transient interactions between an A-domain and CP have been investigated by using acyl vinylsulfonamide adenosine inhibitors as probes to determine the structures of several A-domain-CP complexes. However, this strategy requires a specific vinylsulfonamide inhibitor that contains an acyl group corresponding to the substrate specificity of a target A-domain in every case. Here, we report an alternative strategy for structural characterization of A-domain-CP complexes. We used a bromoacetamide pantetheine cross-linking probe in combination with a Cys mutation to trap the standalone A-domain-CP complex involved in macrolactam polyketide biosynthesis through a covalent linkage, allowing the determination of the complex structure. This strategy facilitates the structural determination of A-domain-CP complexes.

PubMed: 32608966
DOI: 10.1021/acschembio.0c00403

Experimental method

X-RAY DIFFRACTION (2 Å)