Craft John W, Shen Tsai-Wei, Brier Lindsey M, Briggs James M
Department of Biology and Biochemistry, University of Houston , 4800 Calhoun Road, Houston, Texas 77204-5001, United States.
J Phys Chem B. 2015 Jan 22;119(3):1048-61. doi: 10.1021/jp506509c. Epub 2014 Nov 5.
The biophysical chemistry of macromolecular complexes confer their functional characteristics. We investigate the mechanisms that make the AB5 holotoxin of Vibrio cholerae (CT) a significantly more pathogenic molecule than the enterotoxin of Escherichia coli (LT) with which it shares 88% similarity and whose structure is homologous with a backbone RMSD of 0.84 Å and imposes its deleterious effects though the same process to constitutively ADP-ribosylate adenylate cyclase. We present computational data that characterizes the impact of amino acid variations in the A2 tail, which helps to explain experimental data that demonstrate CT's higher toxicity. A hydrophobic patch on the B pentamer interface and its interactions with the A subdomain are partially disrupted by the substitution of an aspartic acid (LT) for glycine in CT. CT's holotoxin has less solvent accessible surface area (94 Å(2) vs 54 Å(2)) and higher contact area (280 Å(2) vs 241 Å(2)) with S228, which is a gatekeeper, partially controlling the diffusion of water into the pore. CT excludes water from the top of the central pore whereas LT allows much more water to interact. These biophysical properties of the toxins lead to their differential toxicity and resulting impact to human health.
大分子复合物的生物物理化学赋予了它们功能特性。我们研究了使霍乱弧菌的AB5全毒素(CT)比大肠杆菌肠毒素(LT)致病性更强的机制,CT与LT有88%的相似性,其结构同源,主链均方根偏差为0.84 Å,且通过相同过程对组成型腺苷酸环化酶进行ADP核糖基化来施加其有害影响。我们展示了计算数据,这些数据表征了A2尾部氨基酸变异的影响,这有助于解释证明CT具有更高毒性的实验数据。在CT中,B五聚体界面上的一个疏水补丁及其与A亚结构域的相互作用因用天冬氨酸(LT)替代甘氨酸而部分受到破坏。CT全毒素与作为守门员的S228的溶剂可及表面积更小(94 Ų对54 Ų),接触面积更大(280 Ų对241 Ų),S228部分控制着水向孔内的扩散。CT将水排除在中央孔顶部,而LT则允许更多的水相互作用。这些毒素的生物物理特性导致了它们不同的毒性以及对人类健康的影响。