7X0F

Structure of Pseudomonas NRPS protein, AmbB-TC bound to Ppant

Summary for 7X0F

• Entry DOI: 10.2210/pdb7x0f/pdb
• Descriptor: AMB antimetabolite synthase AmbB, 4'-PHOSPHOPANTETHEINE (3 entities in total)
• Functional Keywords: non-ribosomal peptide synthetase, ambb, pseudomonas, biosynthetic protein
• Biological source: Pseudomonas aeruginosa PAO1
• Total number of polymer chains: 4
• Total formula weight: 225316.00

Authors

ChuYuanKee, M.,Bharath, S.R.,Song, H. (deposition date: 2022-02-22, release date: 2022-06-15, Last modification date: 2023-11-29)

Primary citation

Chu Yuan Kee, M.J.,Bharath, S.R.,Wee, S.,Bowler, M.W.,Gunaratne, J.,Pan, S.,Zhang, L.,Song, H.
Structural insights into the substrate-bound condensation domains of non-ribosomal peptide synthetase AmbB.
Sci Rep, 12:5353-5353, 2022

PubMed Abstract: Non-ribosomal peptide synthetases (NRPS) are multi-modular/domain enzymes that catalyze the synthesis of bioactive peptides. A crucial step in the process is peptide elongation accomplished by the condensation (C) domain with the aid of a peptidyl carrier or thiolation (T) domain. Here, we examined condensation reaction carried out by NRPS AmbB involved in biosynthesis of L-2-amino-4-methoxy-trans-3-butenoic acid (AMB) in P. aeruginosa. We determined crystal structures of the truncated T-C bidomain of AmbB in three forms, the apo enzyme with disordered T domain, the holo form with serine linked phosphopantetheine (Ppant) and a holo form with substrate (L-alanine) loaded onto Ppant. The two holo forms feature the T domain in a substrate-donation conformation. Mutagenesis combined with functional assays identified residues essential for the attachment of Ppant, anchoring the Ppant-L-Ala in the donor catalytic channel and the role of the conserved His953 in condensation activity. Altogether, these results provide structural insights into the condensation reaction at the donor site with a substrate-bound C domain of AmbB and lay the foundation for understanding the molecular mechanism of condensation which is crucial for AMB synthesis.

PubMed: 35354859
DOI: 10.1038/s41598-022-09188-8

Experimental method

X-RAY DIFFRACTION (2.2 Å)