Wu Xiaolin, Zhang Tao, Mao Mingsong, Zhang Yali, Zhang Zhenpeng, Xu Ping
School of Medicine, Guizhou University, Guiyang, 550025, Guizhou, China.
State Key Laboratory of Proteomics, National Center for Protein Sciences (Beijing), Beijing Proteome Research Center, Beijing Institute of Lifeomics, Beijing, 102206, China.
Proteome Sci. 2024 May 2;22(1):5. doi: 10.1186/s12953-024-00229-w.
Hair is an advantageous biological sample due to its recordable, collectable, and storable nature. Hair's primary components are keratin and keratin-associated proteins. Owing to its abundance of cystine, keratin possesses impressive mechanical strength and chemical stability, formed by creating disulfide bonds as crosslinks within the protein peptide chain. Furthermore, keratin is cross-linked with keratin-associated proteins to create a complex network structure that provides the hair with strength and rigidity. Protein extraction serves as the foundation for hair analysis research. Bleaching hair causes damage to the structure between keratin and keratin-associated proteins, resulting in texture issues and hair breakage. This article outlines various physical treatment methods and lysate analysis that enhance the efficiency of hair protein extraction. The PLEE method achieves a three-fold increase in hair protein extraction efficiency when using a lysis solution containing SDS and combining high temperatures with intense shaking, compared to previous methods found in literature. We utilized the PLEE method to extract hair from both normal and damaged groups. Normal samples identified 156-157 proteins, including 51 keratin and keratin-associated proteins. The damaged group consisted of 155-158 identified proteins, of which 48-50 were keratin and keratin-associated proteins. Bleaching did not cause any notable difference in the protein identification of hair. However, it did reduce coverage of keratin and keratin-associated proteins significantly. Our hair protein extraction method provides extensive coverage of the hair proteome. Our findings indicate that bleaching damage results in subpar hair quality due to reduced coverage of protein primary sequences in keratin and keratin-associated proteins.
由于头发具有可记录、可收集和可储存的特性,它是一种很有优势的生物样本。头发的主要成分是角蛋白和角蛋白相关蛋白。由于富含胱氨酸,角蛋白具有令人印象深刻的机械强度和化学稳定性,它是通过在蛋白质肽链内形成二硫键作为交联来构成的。此外,角蛋白与角蛋白相关蛋白交联形成复杂的网络结构,赋予头发强度和刚性。蛋白质提取是头发分析研究的基础。漂白头发会破坏角蛋白和角蛋白相关蛋白之间的结构,导致发质问题和头发断裂。本文概述了各种提高头发蛋白质提取效率的物理处理方法和裂解物分析。与文献中以前的方法相比,当使用含有十二烷基硫酸钠(SDS)的裂解液并结合高温与剧烈振荡时,PLEE方法可使头发蛋白质提取效率提高三倍。我们利用PLEE方法从正常组和受损组中提取头发。正常样本鉴定出156 - 157种蛋白质,其中包括51种角蛋白和角蛋白相关蛋白。受损组鉴定出155 - 158种蛋白质,其中48 - 50种是角蛋白和角蛋白相关蛋白。漂白对头发蛋白质的鉴定没有造成任何显著差异。然而,它确实显著降低了角蛋白和角蛋白相关蛋白的覆盖率。我们的头发蛋白质提取方法能广泛覆盖头发蛋白质组。我们的研究结果表明,由于角蛋白和角蛋白相关蛋白中蛋白质一级序列的覆盖率降低,漂白损伤导致头发质量不佳。