De novo design of α-amylase inhibitor:: A small linear mimetic of macromolecular proteinaceous ligands
De novo design of α-amylase inhibitor:: A small linear mimetic of macromolecular proteinaceous ligands
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DOI：
10.1016/j.chembiol.2005.10.005
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发表时间：
2005-12-01
影响因子：
--
通讯作者：
Mares, M
中科院分区：
文献类型：
--
作者：
Dolecková-Maresová, L;Pavlík, M;Mares, M
我们报道了一种基于重新设计的线性多肽支架的低分子α-淀粉酶抑制剂，通过组合化学的使用。利用L多肽文库筛选出抑制基序PAMI(多肽淀粉酶抑制因子)，并通过引入非自然修饰进行微调。通过抑制动力学、与天然α-淀粉酶抑制剂的荧光竞争分析和恒温滴定量热法研究了其与猪胰腺α-淀粉酶的相互作用。我们证明了PAMI中的关键氨基酸残基与那些大分子蛋白类抑制剂中的氨基酸残基是相同的，然而，这些大分子蛋白类抑制剂通过一系列分散的分子间接触而与α-淀粉酶结合。因此，自然分子进化和组合进化为完全不同的空间框架选择了相同的α-淀粉酶结合决定因素。
We report a low molecular weight inhibitor of alpha-amylases based on a linear peptidic scaffold designed de novo through the use of combinatorial chemistry. The inhibitory motif denoted PAMI (peptide amylase inhibitor) was selected by using L-peptide libraries and was fine-tuned by the introduction of unnatural modifications. PAMI specifically inhibits glycoside hydrolases of family 13. Its interaction with porcine pancreatic alpha-amylase was characterized by inhibition kinetics, fluorescence competition assays with natural a-amylase inhibitors, and isothermal titration calorimetry. We demonstrate that the critical amino acid residues in PAMI are shared with those in the macromolecular proteinaceous inhibitors that, however, bind to alpha-amylases through a spatially scattered set of intermolecular contacts. Thus, natural molecular evolution as well as combinatorial evolution selected the same alpha-amylase binding determinants for completely different spatial frameworks.