昆虫表皮蛋白OfCPH-1的液液相分离性质

doi:10.16409/j.cnki.2095-039x.2026.01.010

摘要/Abstract

摘要： 昆虫表皮蛋白作为昆虫表皮的主要细胞外基质蛋白，其种类繁多，序列特征多样，参与表皮结构的构建与功能调控，在维持表皮完整性及多样性方面发挥关键作用。亚洲玉米螟Ostrinia furnacalis是我国玉米等作物的主要钻蛀性害虫之一，严重威胁粮食生产安全。前期研究在亚洲玉米螟幼虫头壳中鉴定到一个高丰度CPH家族表皮蛋白OfCPH-1，能够与壳聚糖结合并形成液液相分离（liquid-liquid phase separation,LLPS）凝聚体。本文聚焦于OfCPH-1自身的几丁质结合及LLPS性质研究，以探究其在表皮组装过程中的功能。几丁质结合试验显示，OfCPH-1与α-几丁质有很强的结合能力，尽管其序列中不含有报道的几丁质结合域。相分离性质研究表明，全长OfCPH-1在室温下能够响应离子强度的变化发生LLPS，形成平均粒径为（571.99±8.67）nm的凝聚体，而其截短体多肽18aa和AH-rich基序则不具备相分离能力，这种相分离性质可能与其序列中多个固有无序结构域（IDRs）相关。傅利叶红外光谱（FTIR）分析揭示OfCPH-1发生相分离后，其无规卷曲结构减少而β-折叠结构增加，蛋白从无序状态变得更加有序。本研究为理解CPH家族表皮蛋白的生物学功能提供了新视角，将促进靶向昆虫表皮蛋白相分离过程的新型绿色农药开发。

关键词: 昆虫表皮蛋白, 亚洲玉米螟, 液液相分离

Abstract: Insect cuticular proteins, as the major extracellular matrix components of the insect cuticle, are highly diverse in type and sequence characteristics. They play crucial roles in the construction and functional regulation of the cuticle structure, contributing significantly to the integrity and diversity of the insect exoskeleton. The Asian corn borer Ostrinia furnacalis is one of the main stem-boring pests of maize and other crops in China, posing a serious threat to food security. Previous studies have identified a highly abundant CPH family cuticular protein, OfCPH-1, in the head capsule of O. furnacalis larvae. OfCPH-1 can bind to chitosan and form liquid-liquid phase separation(LLPS) condensates. This study focuses on the chitin-binding ability and LLPS behavior of OfCPH-1 to explore its potential role in cuticle assembly. Chitin-binding assays demonstrated that OfCPH-1 has strong binding affinity to α-chitin, despite lacking any previously reported canonical chitin-binding domains in its sequence. LLPS assays showed that full-length OfCPH-1 undergoes LLPS at room temperature in response to changes in ionic strength, forming condensates with an average diameter of(571.99±8.67) nm, whereas truncated peptides containing the 18 aa motif or AH-rich motif failed to undergo phase separation. The LLPS capability of OfCPH-1 is likely due to the multiple intrinsically disordered regions(IDRs) in its sequence. Fourier-transform infrared spectroscopy(FTIR) analysis further revealed that, upon phase separation, OfCPH-1 undergoes a conformation shift from random coils to β-sheet structures. This study provides new perspective for understanding the biological function of CPH family cuticular proteins and supports the potential development of novel green pesticides by targeting the LLPS behavior of insect cuticular proteins.

Key words: insect cuticular protein, Ostrinia furnacalis, liquid-liquid phase separation

中图分类号:

S476

杜源哲, 陈磊, 杨青. 昆虫表皮蛋白OfCPH-1的液液相分离性质[J]. 中国生物防治学报, 2026, 42(3): 582-590.

DU Yuanzhe, CHEN Lei, YANG Qing. The Liquid-Liquid Phase Separation Properties of the Insect Cuticular Protein OfCPH-1[J]. Chinese Journal of Biological Control, 2026, 42(3): 582-590.

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