Graduate Biomedical Sciences Microbiology Theme, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Acta Crystallogr D Struct Biol. 2017 Nov 1;73(Pt 11):921-939. doi: 10.1107/S2059798317015236. Epub 2017 Oct 26.
Biological chelating molecules called siderophores are used to sequester iron and maintain its ferric state. Bacterial substrate-binding proteins (SBPs) bind iron-siderophore complexes and deliver these complexes to ATP-binding cassette (ABC) transporters for import into the cytoplasm, where the iron can be transferred from the siderophore to catalytic enzymes. In Yersinia pestis, the causative agent of plague, the Yersinia iron-uptake (Yiu) ABC transporter has been shown to improve iron acquisition under iron-chelated conditions. The Yiu transporter has been proposed to be an iron-siderophore transporter; however, the precise siderophore substrate is unknown. Therefore, the precise role of the Yiu transporter in Y. pestis survival remains uncharacterized. To better understand the function of the Yiu transporter, the crystal structure of YiuA (YPO1310/y2875), an SBP which functions to present the iron-siderophore substrate to the transporter for import into the cytoplasm, was determined. The 2.20 and 1.77 Å resolution X-ray crystal structures reveal a basic triad binding motif at the YiuA canonical substrate-binding site, indicative of a metal-chelate binding site. Structural alignment and computational docking studies support the function of YiuA in binding chelated metal. Additionally, YiuA contains two mobile helices, helix 5 and helix 10, that undergo 2-3 Å shifts across crystal forms and demonstrate structural breathing of the c-clamp architecture. The flexibility in both c-clamp lobes suggest that YiuA substrate transfer resembles the Venus flytrap mechanism that has been proposed for other SBPs.
生物螯合分子,称为铁载体,用于螯合铁并维持其三价铁状态。细菌底物结合蛋白 (SBP) 结合铁载体复合物,并将这些复合物递送至 ATP 结合盒 (ABC) 转运体以将其导入细胞质,在此将铁从铁载体转移至催化酶。在鼠疫耶尔森氏菌中,这种鼠疫的病原体,已经表明 Yersinia iron-uptake (Yiu) ABC 转运体在铁螯合条件下改善铁的摄取。该 Yiu 转运体被提议为铁载体转运体;然而,确切的铁载体底物是未知的。因此,Yiu 转运体在鼠疫耶尔森氏菌存活中的确切作用仍未被阐明。为了更好地理解 Yiu 转运体的功能,测定了 YiuA(YPO1310/y2875)的晶体结构,YiuA 是一种 SBP,其功能是将铁载体底物呈现给转运体以将其导入细胞质。2.20 和 1.77 Å 的 X 射线晶体结构揭示了 YiuA 经典底物结合位点处的基本三联体结合基序,表明了金属螯合物结合位点。结构比对和计算对接研究支持 YiuA 在结合螯合金属中的功能。此外,YiuA 包含两个可移动的螺旋,即螺旋 5 和螺旋 10,它们在晶体形式之间发生 2-3 Å 的位移,并表现出 c-夹结构的呼吸。两个 c-夹叶的柔韧性表明,YiuA 底物的转移类似于已被提议用于其他 SBP 的 Venus flytrap 机制。