Crystallographic analysis of NosA, which catalyzes terminal amide formation in the biosynthesis of nosiheptide.

Wang, Yanting; Liu, Shanshan; Yao, Pengfei; Yu, Yi; Zhang, Yan; Lan, Wenxian; Wang, Chunxi; Ding, Jiuping; Liu, Wen; Cao, Chunyang

Nosiheptide is a member of the thiopeptide family of antibiotics which demonstrates potent activities against various bacterial pathogens. The formation of its C-terminal amide is catalysed by NosA in an unusual strategy for maturating certain thiopeptides by processing precursor peptides featuring a serine extension. Here, a recombinant C-terminally truncated selenomethionine-derivatized NosA1-111 variant from Streptomyces actuosus consisting of residues 1-111, named SeMet NosA1-111, was crystallized using the sitting-drop vapour-diffusion method. Diffraction data were collected to 2.40 Å resolution using synchrotron radiation. The crystals belonged to the primitive cubic space group P4132, with unit-cell parameters a = b = c = 143.3 Å. Assuming the presence of three molecules in the asymmetric unit, the calculated Matthews coefficient was 3.94 Å3 Da−1 and the corresponding solvent content was 40.3%.

THIOPEPTIDES; ANTIBIOTIC synthesis; PEPTIDE antibiotics; CRYSTALLOGRAPHY; AMIDE synthesis; AMIDES; CATALYTIC activity; CRYSTAL structure

Acta Crystallographica: Section F, Structural Biology Communications, 2015, Vol 71, Issue 8, p1033

2053-230X

Academic Journal

10.1107/S2053230X15011085