Interaction of reduced nicotinamide adenine dinucleotide with an antifreeze protein from Dendroides canadensis: mechanistic implication of antifreeze activity enhancement.
Interaction of reduced nicotinamide adenine dinucleotide with an antifreeze protein from Dendroides canadensis: mechanistic implication of antifreeze activity enhancement.
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DOI：
10.1002/jmr.1151
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发表时间：
2011-11
期刊：
影响因子：
--
通讯作者：
Sacco MA
中科院分区：
文献类型：
--
作者：
Wen X;Wang S;Amornwittawat N;Houghton EA;Sacco MA
抗冻蛋白（AFP）在许多生物体中发现，可以非聚集性地降低水的冰点，而不改变熔点。降低的凝固点和熔点之间的差异，称为热滞后（TH），通常是AFP抗冻活性的量度。某些低分子量的分子和蛋白质可以进一步增强AFP的抗冻活性。增强子和精氨酸之间的相互作用在增强加拿大石斛甲胎蛋白（DAFP-1）的抗冻活性中起重要作用。在这里，我们研究了几种常见的含磷酸辅酶对DAFP-1抗冻活性的增强作用。β-还原型烟酰胺腺嘌呤二核苷酸（NADH）被认为是DAFP-1最有效的增强子，可使DAFP-1的抗冻活性提高约10倍。对一系列NADH类似物和NADH的各种分子片段的增强能力的检查表明，烟酰胺的修饰产生了一系列高效的增强剂，尽管没有像NADH本身那样有效，并且NADH的整个分子结构是其高效增强效应所必需的。我们还证明了DAFP-1和NADH之间的1：1结合。通过使用Hummel和Dreyer的凝胶过滤方法的高效液相色谱（HPLC）表征结合。数据分析表明DAFP-1和NADH之间的结合具有微摩尔范围内的解离常数。DAFP-1和NADH之间的相互作用与增强子作用的分子机制一起讨论沿着。
Antifreeze proteins (AFPs) found in many organisms can noncolligatively lower the freezing point of water without altering the melting point. The difference between the depressed freezing point and the melting point, termed thermal hysteresis (TH), is usually a measure of the antifreeze activity of AFPs. Certain low molecular mass molecules and proteins can further enhance the antifreeze activity of AFPs. Interaction between an enhancer and arginine is known to play an important role in enhancing the antifreeze activity of an AFP from the beetle Dendroides canadensis (DAFP-1). Here, we examined the enhancement effects of several prevalent phosphate-containing coenzymes on the antifreeze activity of DAFP-1. β-Nicotinamide adenine dinucleotide (reduced) (NADH) is identified as the most efficient enhancer of DAFP-1, which increases the antifreeze activity of DAFP-1 by around 10 times. Examination of the enhancement abilities of a series of NADH analogs and various molecular fragments of NADH reveals that the modifications of nicotinamide generate a series of highly efficient enhancers, though none as effective as NADH itself, and the whole molecular structure of NADH is necessary for its highly efficient enhancement effect. We also demonstrated a 1:1 binding between DAFP-1 and NADH. The binding was characterized by high-performance liquid chromatography (HPLC) using the gel filtration method of Hummel and Dreyer. The data analysis suggests binding between DAFP-1 and NADH with a dissociation constant in the micromolar range. Interactions between DAFP-1 and NADH are discussed along with molecular mechanisms of enhancer action.