3GZM

Crystal Structure of holo PfACP Reduced Monomer

3GZM の概要

• エントリーDOI: 10.2210/pdb3gzm/pdb
• 関連するPDBエントリー: 3GZL
• 分子名称: Acyl carrier protein, 4'-PHOSPHOPANTETHEINE, BETA-MERCAPTOETHANOL, ... (5 entities in total)
• 機能のキーワード: helix bundle, phosphopantetheine, fatty acid biosynthesis, lipid synthesis, transit peptide, biosynthetic protein
• 由来する生物種: Plasmodium falciparum
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 20604.12

構造登録者

Gallagher, J.R.,Prigge, S.T. (登録日: 2009-04-07, 公開日: 2009-04-21, 最終更新日: 2023-09-06)

主引用文献

Gallagher, J.R.,Prigge, S.T.
Plasmodium falciparum acyl carrier protein crystal structures in disulfide-linked and reduced states and their prevalence during blood stage growth.
Proteins, 78:575-588, 2010

PubMed Abstract: Acyl Carrier Protein (ACP) has a single reactive sulfhydryl necessary for function in covalently binding nascent fatty acids during biosynthesis. In Plasmodium falciparum, the causative agent of the most lethal form of malaria, fatty acid biosynthesis occurs in the apicoplast organelle during the liver stage of the parasite life cycle. During the blood stage, fatty acid biosynthesis is inactive and the redox state of the apicoplast has not been determined. We solved the crystal structure of ACP from P. falciparum in reduced and disulfide-linked forms, and observe the surprising result that the disulfide in the PfACP cross-linked dimer is sequestered from bulk solvent in a tight molecular interface. We assessed solvent accessibility of the disulfide with small molecule reducing agents and found that the disulfide is protected from BME but less so for other common reducing agents. We examined cultured P. falciparum parasites to determine which form of PfACP is prevalent during the blood stages. We readily detected monomeric PfACP in parasite lysate, but do not observe the disulfide-linked form, even under conditions of oxidative stress. To demonstrate that PfACP contains a free sulfhydryl and is not acylated or in the apo state, we treated blood stage parasites with the disulfide forming reagent diamide. We found that the effects of diamide are reversed with reducing agent. Together, these results suggest that the apicoplast is a reducing compartment, as suggested by models of P. falciparum metabolism, and that PfACP is maintained in a reduced state during blood stage growth.

PubMed: 19768685
DOI: 10.1002/prot.22582

実験手法

X-RAY DIFFRACTION (1.8 Å)