6TG5

Solution structure of MacpD, a acyl carrier protein, from Pseudomonas fluorescens involved in Mupirocin biosynthesis.

Summary for 6TG5

• Entry DOI: 10.2210/pdb6tg5/pdb
• NMR Information: BMRB: 34451
• Descriptor: MacpD (1 entity in total)
• Functional Keywords: mupirocin, acyl carrier protein, acyl starter units, thiomarinol, biosynthesis, pseudomonas fluorescens, biosynthetic protein
• Biological source: Pseudomonas fluorescens
• Total number of polymer chains: 1
• Total formula weight: 14936.70

Authors

Williams, C.,Crump, M.P. (deposition date: 2019-11-15, release date: 2020-02-19, Last modification date: 2024-06-19)

Primary citation

Walker, P.D.,Rowe, M.T.,Winter, A.J.,Weir, A.N.M.,Akter, N.,Wang, L.,Race, P.R.,Williams, C.,Song, Z.,Simpson, T.J.,Willis, C.L.,Crump, M.P.
A Priming Cassette Generates Hydroxylated Acyl Starter Units in Mupirocin and Thiomarinol Biosynthesis.
Acs Chem.Biol., 15:494-503, 2020

PubMed Abstract: Mupirocin, a commercially available antibiotic produced by NCIMB 10586, and thiomarinol, isolated from the marine bacterium sp. SANK 73390, both consist of a polyketide-derived monic acid homologue esterified with either 9-hydroxynonanoic acid (mupirocin, 9HN) or 8-hydroxyoctanoic acid (thiomarinol, 8HO). The mechanisms of formation of these deceptively simple 9HN and 8HO fatty acid moieties in and , respectively, remain unresolved. To define starter unit generation, the purified mupirocin proteins MupQ, MupS, and MacpD and their thiomarinol equivalents (TmlQ, TmlS and TacpD) have been expressed and shown to convert malonyl coenzyme A (CoA) and succinyl CoA to 3-hydroxypropionoyl (3-HP) or 4-hydroxybutyryl (4-HB) fatty acid starter units, respectively, the MupQ/TmlQ catalyzed generation of an unusual bis-CoA/acyl carrier protein (ACP) thioester, followed by MupS/TmlS catalyzed reduction. Mix and match experiments show MupQ/TmlQ to be highly selective for the correct CoA. MacpD/TacpD were interchangeable but alternate -acting ACPs from the mupirocin pathway (MacpA/TacpA) or a heterologous ACP (BatA) were nonfunctional. MupS and TmlS selectivity was more varied, and these reductases differed in their substrate and ACP selectivity. The solution structure of MacpD determined by NMR revealed a C-terminal extension with partial helical character that has been shown to be important for maintaining high titers of mupirocin. We generated a truncated MacpD construct, MacpD_T, which lacks this C-terminal extension but retains an ability to generate 3-HP with MupS and MupQ, suggesting further downstream roles in protein-protein interactions for this region of the ACP.

PubMed: 31977176
DOI: 10.1021/acschembio.9b00969

Experimental method

SOLUTION NMR