Liu Weizhi, Du Li, Zhang Liang, Chen Jing, Shen Xu, Jiang Hualiang
Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Shanghai 201203, China.
Protein Expr Purif. 2007 Mar;52(1):74-81. doi: 10.1016/j.pep.2006.09.003. Epub 2006 Sep 15.
Acyl carrier protein (ACP) is an essential component in the type II fatty acid biosynthesis (FAS II) process and is responsible for the acyl group transfer within a series of related enzymes. In this work, the ACP from Helicobacter pylori strain SS1 was cloned and the gene sequence of Hpacp was deposited in the GenBank database (Accession No.: AY904356). Two forms of HpACP (apo, holo) were successfully purified and characterized. The thermal stability of these two forms was quantitatively investigated by CD spectral analyses. The results revealed that the holo-HpACP was more stable than apo-HpACP according to the transition midpoint temperature(Tm). Moreover, the interaction of HpACP with the related enzyme (beta-hydroxyacyl-ACP dehydratase, HpFabZ) was determined by GST-pull down assay and surface plasmon resonance (SPR) technique in vitro, the results showed that HpACP displays a strong binding affinity to HpFabZ (KD=1.2 x 10(-8)M). This current work is hoped to supply useful information for better understanding the ACP features of Helicobacter pylori SS1 strain.
酰基载体蛋白（ACP）是II型脂肪酸生物合成（FAS II）过程中的一个重要组成部分，负责在一系列相关酶中进行酰基转移。在本研究中，克隆了幽门螺杆菌SS1菌株的ACP，并将Hpacp的基因序列存入GenBank数据库（登录号：AY904356）。成功纯化并鉴定了两种形式的HpACP（脱辅基蛋白、全蛋白）。通过圆二色光谱分析对这两种形式的热稳定性进行了定量研究。结果表明，根据转变中点温度（Tm），全蛋白-HpACP比脱辅基蛋白-HpACP更稳定。此外，通过体外GST下拉试验和表面等离子体共振（SPR）技术测定了HpACP与相关酶（β-羟基酰基-ACP脱水酶，HpFabZ）的相互作用，结果表明HpACP对HpFabZ表现出很强的结合亲和力（KD=1.2×10^(-8)M）。希望这项工作能为更好地了解幽门螺杆菌SS1菌株的ACP特性提供有用信息。
