人维生素K环氧化物还原酶复合物(hVKORC1),一种将维生素K转化为血液凝固所需形式的关键酶,需要其氧化还原伙伴通过硫醇-二硫化物交换反应递送的还原当量来活化。腔环路(L-环路)是hVKORC1激活的主要介质,这是一个经常藏有许多错义突变的地区。四个L环hVKORC1突变体,在体外建议为抗性(A41S,H68Y)或完全不工作(S52W,W59R),通过数值方法(计算机模拟)在氧化状态下进行了研究。对每个突变体的DYNASOME和POCKETOME进行了表征,并与天然蛋白质进行了比较。最近被描述为由结构稳定的跨膜结构域(TMD)和固有无序的L环组成的模块蛋白,表现出准独立的动力学。突变体的DYNASOME表明,L-loop错义点突变不仅影响其折叠和动力学,还有TMD的,突出了这些结构域之间强烈的突变特异性相互依存。突变诱导效应的另一个结果体现在全局变化(几何,拓扑,和概率)新检测到的隐蔽袋以及L环与其氧化还原蛋白的识别特性的交替。根据我们的结果,我们假设(i)蛋白质内变构调节,(ii)每个突变体的固有变构调节和隐伏袋取决于其DYNASOME;(iii)hVKORC1(INTERACTOME)对氧化还原蛋白的识别取决于其DYNASOME。这种对蛋白质的多方面描述产生了“组学”数据集,对于理解蛋白质的生理过程和由各种“组学”水平的蛋白质性质改变引起的病理至关重要。此外,这种特征为血液疾病治疗所必需的“全网络药物”的开发开辟了新的前景。
The human Vitamin K Epoxide Reductase Complex (hVKORC1), a key enzyme transforming vitamin K into the form necessary for blood clotting, requires for its activation the reducing equivalents delivered by its redox partner through thiol-disulfide exchange reactions. The luminal loop (L-loop) is the principal mediator of hVKORC1 activation, and it is a region frequently harbouring numerous missense mutations. Four L-loop hVKORC1 mutants, suggested in vitro as either resistant (A41S, H68Y) or completely inactive (S52W, W59R), were studied in the oxidised state by numerical approaches (in silico). The DYNASOME and POCKETOME of each mutant were characterised and compared to the native protein, recently described as a modular protein composed of the structurally stable transmembrane domain (TMD) and the intrinsically disordered L-loop, exhibiting quasi-independent dynamics. The DYNASOME of mutants revealed that L-loop missense point mutations impact not only its folding and dynamics, but also those of the TMD, highlighting a strong mutation-specific interdependence between these domains. Another consequence of the mutation-induced effects manifests in the global changes (geometric, topological, and probabilistic) of the newly detected cryptic pockets and the alternation of the recognition properties of the L-loop with its redox protein. Based on our results, we postulate that (i) intra-protein allosteric regulation and (ii) the inherent allosteric regulation and cryptic pockets of each mutant depend on its DYNASOME; and (iii) the recognition of the redox protein by hVKORC1 (INTERACTOME) depend on their DYNASOME. This multifaceted description of proteins produces \"omics\" data sets, crucial for understanding the physiological processes of proteins and the pathologies caused by alteration of the protein properties at various \"omics\" levels. Additionally, such characterisation opens novel perspectives for the development of \"allo-network drugs\" essential for the treatment of blood disorders.