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Crystallization and X‐ray analysis of Borrelia burgdorferi β‐barrel assembly machinery A
Author(s) -
Dong Shishang,
Chu Hongguan,
Wen Kangning,
Yu Qianqian,
Li Hui,
Wang Changhui,
Qin Xiaochun
Publication year - 2020
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x20006196
Subject(s) - bama , periplasmic space , barrel (horology) , crystallization , borrelia burgdorferi , biology , eukaryote , bacterial outer membrane , snare complex , biophysics , membrane , crystallography , chemistry , biochemistry , materials science , genome , escherichia coli , gene , genetics , vesicle , antibody , organic chemistry , composite material
Mitochondria, chloroplasts and several species of bacteria have outer membrane proteins (OMPs) that perform many essential biological functions. The β‐barrel assembly machinery (BAM) complex is one of the OMPs of Borrelia burgdorferi , the pathogenic spirochete that causes Lyme disease, and its BamA component ( Bb BamA) includes a C‐terminal β‐barrel domain and five N‐terminal periplasmic polypeptide‐transport‐associated (POTRA) domains, which together perform a central transport function. In the current work, the production, crystallization and X‐ray analysis of the three N‐terminal POTRA domains of Bb BamA ( Bb BamA‐POTRA P1–P3; residues 30–273) were carried out. The crystals of Bb BamA‐POTRA P1–P3 belonged to space group P 2 1 , with unit‐cell parameters a = 45.353, b = 111.538, c = 64.376 Å, β = 99.913°. The Matthews coefficient was calculated to be 2.92 Å 3 Da −1 , assuming the presence of two molecules per asymmetric unit, and the corresponding solvent content was 57.9%. Owing to the absence of an ideal homology model, numerous attempts to solve the Bb BamA‐POTRA P1–P3 structure using molecular replacement (MR) failed. In order to solve the structure, further trials using selenomethionine derivatization are currently being carried out.