Santos Lucianna Helene S, Broilo Campos Augusto César, Corrêa Santos Viviane, Fassio Alexandre Victor, Costa Maurício G S, Ferreira Rafaela Salgado
Institut Pasteur de Montevideo, Mataojo 2020, Montevideo 11400, Uruguay.
Departamento de Bioquímica e Imunologia, Universidade Federal de Minas Gerais, Avenida Antônio Carlos 6627, Belo Horizonte 31270-901, Minas Gerais, Brazil.
ACS Omega. 2025 May 2;10(18):19115-19128. doi: 10.1021/acsomega.5c01876. eCollection 2025 May 13.
Chagas disease (CD) is a neglected tropical disease for which novel and improved treatments are needed. The cysteine protease Cruzipain is one of the main targets for the development of novel drugs for the treatment of CD. Recent bioinformatics analyses have revealed four Cruzipain subtypes whose active sites differ in key positions for ligand recognition. These analyses suggest a possible effect on the substrate specificity and affinity for ligands. To better investigate the impact of substitutions in Cruzipain subtypes, we employed molecular dynamics simulations and varied structural analyses on representatives of each Cruzipain subtype. Our results indicated that the substitutions did not significantly affect the overall flexibility and conformation of these proteases. In contrast, we observed differences in their active site characteristics, including different electrostatic potentials, cavity volumes, and patterns of interactions with the virtual probes. The distinct alterations in the active site subsites, especially in the S2 subsite, suggest unique functional changes that could affect substrate binding, ligand recognition, and possibly enzymatic effectiveness in various biological situations.
恰加斯病（CD）是一种被忽视的热带疾病，需要新颖且改良的治疗方法。半胱氨酸蛋白酶克氏锥虫蛋白酶是开发治疗CD新药的主要靶点之一。最近的生物信息学分析揭示了四种克氏锥虫蛋白酶亚型，其活性位点在配体识别的关键位置上有所不同。这些分析表明对底物特异性和配体亲和力可能有影响。为了更好地研究克氏锥虫蛋白酶亚型中取代的影响，我们对每种克氏锥虫蛋白酶亚型的代表进行了分子动力学模拟和各种结构分析。我们的结果表明，这些取代并未显著影响这些蛋白酶的整体灵活性和构象。相反，我们观察到它们活性位点特征的差异，包括不同的静电势、腔体积以及与虚拟探针的相互作用模式。活性位点亚位点的明显改变，特别是在S2亚位点，表明可能存在独特的功能变化，这可能会影响底物结合、配体识别，并可能在各种生物学情况下影响酶的有效性。
