蛋白质翻译后修饰（post-translation modifications，PTMs）是发生于蛋白质生物合成中的化学修饰，与蛋白质功能及参与各种生命活动密切相关，如DNA的复制和转录，蛋白质翻译，细胞的信号转导和通讯等。赖氨酸巴豆酰化是近年来新发现的一种PTM，在各种重要的调节途径中起关键作用。红色毛癣菌作为研究浅表感染和丝状真菌的模式生物，被广泛用于真菌的致病和耐药机制的研究。本文基于巴豆酰化肽段的富集和高通量LC-MS/MS（Liquid Chromatography Tandem Mass Spectrometry）检测，对红色毛癣菌的两个主要生长阶段—孢子和菌丝的巴豆酰化修饰进行了全蛋白质组水平的鉴定；研究巴豆酰化蛋白在红色毛癣菌中的亚细胞定位和功能，并且通过对比菌丝时相和孢子时相巴豆酰化修饰的差异，对巴豆酰化蛋白参与红色毛癣菌的基因表达和生长发育调控进行初步研究。

在红色毛癣菌孢子和菌丝时相中共鉴定出3144个巴豆酰化蛋白以及14019个巴豆酰化位点，占整个红色毛癣菌蛋白质组的30%。此外，巴豆酰化修饰具有较高的蛋白修饰位点率，平均每个巴豆酰化蛋白约有4.45个巴豆酰化位点。在孢子时相鉴定到2764个蛋白上的11300个巴豆酰化位点；在菌丝时相鉴定到1954个蛋白上的8230个巴豆酰化位点。其中1574个蛋白在两个时相均发生巴豆酰化修饰，且存在5511个共有的巴豆酰化位点；1190个蛋白在孢子阶段特异性修饰，380个蛋白在菌丝阶段特异性修饰。对巴豆酰化蛋白进行GO和KEGG分析发现，巴豆酰化蛋白广泛参与各种生物合成和代谢途径，这些结果说明巴豆酰化修饰可能参与到红色毛癣菌不同生长阶段的调控中。

组蛋白的翻译后修饰是表观遗传的重要机制。本文在组蛋白上共鉴定到53个巴豆酰化修饰位点。其中34个巴豆酰化位点在孢子和菌丝时相共同存在；17个巴豆酰化位点特异修饰于孢子阶段，2个位点特异修饰于菌丝阶段。此外，45种分泌蛋白酶和30种外排转运蛋白中鉴定存在巴豆酰化修饰。由于分泌蛋白是红色毛癣菌关键的毒力因子，而外排转运蛋白在耐药过程中起到重要作用，这些结果表明，巴豆酰化不仅可能广泛参与各种生物合成和代谢途径，同时也在红色毛癣菌感染和耐药方面发挥调控作用。

本论文首次在蛋白质组水平鉴定了红色毛癣菌的巴豆酰化修饰，并对比了真菌不同生长阶段巴豆酰化修饰的差异，对巴豆酰化蛋白功能、亚细胞定位及巴豆酰化修饰位点特征等进行了系统研究。研究结果也为其他的浅表感染真菌的蛋白质翻译后修饰的研究奠定了基础。

Protein post-translational modifications (PTMs) are chemical alterations that occur subsequent to protein biosynthesis, playing a pivotal role in modulating protein functionality and extensively contributing to a range of biological processes, including DNA replication and transcription, protein translation, and cellular signal transduction and communication. Among these modifications, lysine crotonylation, a recently identified PTM, has gained recognition as a significant regulatory modification implicated in several critical biological pathways. Trichophyton rubrum, a model organism for studying superficial fungal infections, has been extensively employed to investigate fungal pathogenicity and mechanisms of drug resistance. In this study, we conducted an extensive proteome-wide analysis to identify lysine crotonylation in T. rubrum during two critical growth stages: conidia and mycelia. This was achieved through the enrichment of crotonylated peptides in conjunction with high-throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Furthermore, we examined the subcellular localization and functional characteristics of crotonylated proteins, comparing the differential crotonylation profiles between the conidia and mycelia stages. This comparison aimed to elucidate the role of crotonylated proteins in the regulation of gene expression and the control of developmental growth processes. A comprehensive analysis identified 3,144 crotonylated proteins encompassing 14,019 crotonylation sites across both developmental stages, accounting for approximately 30% of the total T. rubrum proteome. Notably, the modification of crotonylation was prevalent, with an average of 4.45 sites per crotonylated protein, underscoring its abundant presence within T. rubrum. In the conidia stage, a total of 11,300 crotonylation sites were identified across 2,764 proteins, whereas the mycelia stage exhibited 8,230 sites across 1,954 proteins. Notably, 1,574 proteins containing 5,511 common crotonylation sites were present in both stages. Additionally, 1,190 proteins were uniquely modified in the conidia, and 380 proteins were uniquely modified in the mycelia. These findings indicate that differential crotonylation plays a role in regulating the distinct growth stages of T. rubrum. Further analysis using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) demonstrated that crotonylated proteins are significantly involved in a wide array of biosynthetic and metabolic pathways. Histone post-translational modifications are fundamental epigenetic regulatory mechanisms. In this study, 53 histone crotonylation sites were identified, with 34 sites common to both conida and mycelia, 17 unique toconidia, and 2 specific to mycelia. Furthermore, crotonylation modifications were observed in 45 secreted proteases and 30 efflux transporter proteins. Considering that secreted proteins are crucial virulence factors in T. rubrum infections and efflux transporters are key to drug resistance, these findings indicate that crotonylation significantly influences various biosynthetic and metabolic pathways, playing a critical role in the regulation of infection and drug resistance in T. rubrum. This dissertation presents the first proteome-wide characterization of crotonylation in T. rubrum, systematically comparing crotonylation profiles across fungal developmental stages and thoroughly analyzing the functional roles, subcellular localization, and site-specific characteristics of crotonylated proteins. These results provide foundational insights for future research into post-translational modifications of other superficial pathogenic fungi.