当前基因组测序的能力超过了功能解释。我们之前的工作表明，3D 蛋白质结构计算增强了对测序肿瘤和罕见疾病患者遗传变异的机制理解。 KRAS GTPase 是驱动癌症和种系疾病的关键遗传因素之一。由于 KRAS 改变的肿瘤经常含有三种经典热点突变之一，因此几乎所有研究都集中在这些突变上，从而在癌症和非癌症疾病中观察到的更广泛的 KRAS 基因组景观中留下了显着的功能模糊性。在此，我们通过分子模拟扩展结构生物信息学，以研究 86 个 KRAS 突变的扩展景观。我们确定了与实验建立的 KRAS 生物物理和生化特性密切相关的多种协调变化。我们观察到的模式跨越热点和非热点改变，这些改变都可以使开关区域失调，产生具有不同效应子结合倾向的突变限制构象。我们通过实验测量了突变热稳定性，并通过模拟识别了共享和不同的模式。我们的结果表明了突变特异性构象，这显示了未来研究这些改变如何影响不同分子和细胞功能的潜力。我们提供的数据无法使用当前的基因组工具进行预测，这证明了从分子模拟中获得的附加功能信息，用于解释人类遗传变异。© 2023 作者。

Current capabilities in genomic sequencing outpace functional interpretations. Our previous work showed that 3D protein structure calculations enhance mechanistic understanding of genetic variation in sequenced tumors and patients with rare diseases. The KRAS GTPase is among the critical genetic factors driving cancer and germline conditions. Because KRAS-altered tumors frequently harbor one of three classic hotspot mutations, nearly all studies have focused on these mutations, leaving significant functional ambiguity across the broader KRAS genomic landscape observed in cancer and non-cancer diseases. Herein, we extend structural bioinformatics with molecular simulations to study an expanded landscape of 86 KRAS mutations. We identify multiple coordinated changes strongly associated with experimentally established KRAS biophysical and biochemical properties. The patterns we observe span hotspot and non-hotspot alterations, which can all dysregulate Switch regions, producing mutation-restricted conformations with different effector binding propensities. We experimentally measured mutation thermostability and identified shared and distinct patterns with simulations. Our results indicate mutation-specific conformations, which show potential for future research into how these alterations reverberate into different molecular and cellular functions. The data we present is not predictable using current genomic tools, demonstrating the added functional information derived from molecular simulations for interpreting human genetic variation.© 2023 The Authors.