PROBING CELLULAR INTRACELLULAR CALCIUM SIGNALING AND SENSING THROUGH COMPUTATION

2+ 细胞内钙信号Kekenes-Huskey，PM肯塔基大学 细胞内钙信号转导机制的研究 计算 钙信号调节广泛的组织类型和物种的生物功能， 2+ 但已知的几个控制钙依赖信号转导和fi活性的因素对这两种计算机都构成了挑战。 分析性的和实验性的探究。在我们对fi的细微差别的理解上有明显的差距 蛋白质的结构和动力学以及它们在细胞内的分布对根本上重要 2+ 过程包括1)钙如何在局部胞内区积累2)蛋白质结合 2+2+ 具有高affi的CA 3)Ca‘感受器蛋白调节信号级联。详细了解以下内容 这些主题及其相互依存关系将产生我们如何看待生物学、生理学、 和健康。在这方面，计算机模拟是有吸引力的，这两个方面都描述了 DIFfiCUT直接通过实验解决，以及形成跨 这些基本主题。然而，有几个突出的障碍导致了这种变革性的模拟 成本令人望而却步。其中，减少了与模型相关的棘手的计算开销- INGfiNe详细描述了由亚纳米到微米尺度、原子尺度 影响离子/蛋白质结合的热力学因素，以及促进蛋白质/蛋白质结合的长程力 信号通路，可能是当今生物物理学面临的最大挑战。在这份提案中，我们概述了几个 外设的多尺度算法进步将缓解这一挑战，同时提供对重要的 2+ CA驱动的流程可协调生活： 主题1在分子水平上调整钙离子的感知和响应。在这个主题中，我们将 开发新的范例来理解自然控制特定fi城市和动力学的诀窍 2+ CA传感功能。 主题2心脏疾病相关形态变化的自动检测 2+ 细胞及其fl对钙稳态的影响。在这个主题中，我们的实验室将利用 未充分利用显微镜数据来回答有关细胞内组织的作用的问题 2+ 在塑造钙信号方面。 主题3：通过P2X4受体控制细胞尺度的分子机制。在这 主题，我们将在分子尺度的蛋白质结构/功能和它们的 控制细胞尺度的信号结果。 1

2+ Intracellular Ca signaling Kekenes-Huskey, PM. University of Kentucky Probing cellular intracellular calcium signaling and sensing through computation Calcium signaling regulates biological function across a broad range of tissue types and species, 2+ but several factors known to control Ca -dependent signaling efficiency have challenged both compu- tational and experimental inquiry. There are significant gaps in our understanding of how nuances in protein structure and dynamics as well as their intracellular distribution affect fundamentally important 2+ processes including how 1) Ca accumulates within localized intracellular regions 2) proteins bind 2+ 2+ Ca with high affinity 3) Ca 'sensor' proteins regulate signaling cascades. Detailed knowledge about these topics and their inter-dependencies would yield new paradigms in how we view biology, physiology, and health. Computer simulations are attractive in this regard, both for describing phenomena that are difficult to directly resolve experimentally, as well as forming integrative conceptual models spanning these underlying topics. However, several prominent hurdles render such transformative simulations cost-prohibitive. Among these, reducing the intractable computational expense involved with model- ing fine detail processes like transport governed by sub-nanometer to micron scales, atomistic-scale thermodynamic factors shaping ion/protein binding, and long-range forces that promote protein/protein signaling pathways, is likely the foremost challenge in biophysics today. In this proposal, we outline sev- eral multi-scale algorithmic advances that will ease this challenge, while providing insight into important 2+ Ca -driven processes that orchestrate life: Theme 1 Tuning Ca2+ sensing and response at the molecular level. In this theme, we will develop new paradigms for understanding nature's tricks for controlling specificity and kinetics in 2+ Ca sensing functions. Theme 2 Automated detection of disease-associated morphological changes in cardiac 2+ cells and their influence on Ca homeostasis. In this theme, our lab will leverage troves of underutilized microscopy data to answer questions regarding the role of intracellular organization 2+ in shaping Ca signaling. Theme 3 Molecular mechanisms of cellular-scale control via the P2X4 receptor. In this theme, we will establish strong links between molecular scale protein structure/function and their control of cellular-scale signaling outcomes. 1