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Exactly how the missense mutations in the GAP\u2010related domain of NF1 (NF1<jats:sup>GRD<\/jats:sup>) allosterically impact NF1 GAP to promote these distinct pathologies is unclear. Especially tantalizing is the question of how same\u2010domain, same\u2010residue NF1<jats:sup>GRD<\/jats:sup> variants exhibit distinct clinical phenotypes. Guided by clinical data, we take up this dilemma. We sampled the conformational ensembles of NF1<jats:sup>GRD<\/jats:sup> in complex with GTP\u2010bound K\u2010Ras4B by performing molecular dynamics simulations. Our results show that mutations in NF1<jats:sup>GRD<\/jats:sup> retain the active conformation of K\u2010Ras4B but with biased propensities of the catalytically competent populations of K\u2010Ras4B\u2013NF1<jats:sup>GRD<\/jats:sup> complex. In agreement with clinical depiction and experimental tagging, compared to the wild type, NF1<jats:sup>GRD<\/jats:sup> E1356A and E1356V mutants effectively act through loss\u2010of\u2010function and gain\u2010of\u2010function mechanisms, leading to neurofibromatosis and developmental disorders, respectively. Allosteric modulation of NF1<jats:sup>GRD<\/jats:sup> GAP activity through biasing the conformational ensembles in the different states is further demonstrated by the diminished GAP activity by NF1<jats:sup>GRD<\/jats:sup> isoform 2, further manifesting propensities of conformational ensembles as powerful predictors of protein function. Taken together, our work identifies a NF1<jats:sup>GRD<\/jats:sup> hotspot that could allosterically tune GAP function, suggests targeting Ras oncogenic mutations by restoring NF1 catalytic activity, and offers a molecular mechanism for NF1 phenotypes determined by their distinct conformational propensities.<\/jats:p>","DOI":"10.1002\/pro.70042","type":"journal-article","created":{"date-parts":[[2025,1,22]],"date-time":"2025-01-22T12:30:52Z","timestamp":1737549052000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Allosteric modulation of <scp>NF1 GAP<\/scp>: Differential distributions of catalytically competent populations in loss\u2010of\u2010function and gain\u2010of\u2010function mutants"],"prefix":"10.1002","volume":"34","author":[{"given":"Liang","family":"Xu","sequence":"first","affiliation":[{"name":"Computational Structural Biology Section, Frederick National Laboratory for Cancer Research in the Cancer 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