Uranga Carla C, Ghassemian Majid, Hernández-Martínez Rufina
Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE), Carretera Ensenada-Tijuana 3918, Zona Playitas, 22860 Ensenada, B.C., Mexico.
University of California, San Diego, Department of Chemistry and Biochemistry, 9500 Gilman Drive, La Jolla, CA 92093-0378, USA.
Biochim Open. 2017 Mar 14;4:88-98. doi: 10.1016/j.biopen.2017.03.001. eCollection 2017 Jun.
Many basic science questions remain regarding protein functions in the pathogen: host interaction, especially in the trunk disease fungi family, the Botryosphaeriaceae, which are a global problem for economically important plants, especially fruiting trees. Proteomics is a highly useful technology for studying protein expression and for discovering novel proteins in unsequenced and poorly annotated organisms. Current fungal proteomics approaches involve 2D SDS-PAGE and extensive, complex, protein extraction methodologies. In this work, a modified Folch extraction was applied to protein extraction to perform both peptide sequencing and peptide fragmentation analysis/protein identification of the plant and human fungal pathogen . Both bioinformatics approaches yielded novel peptide sequences from proteins produced by in the presence of exogenous triglycerides and glucose. These proteins and the functions they may possess could be targeted for further functional characterization and validation efforts, due to their potential uses in biotechnology and as new paradigms for understanding fungal biochemistry, such as the finding of allergenic enolases, as well as various novel proteases, including zinc metalloproteinases homologous to those found in snake venom. This work contributes to genomic annotation efforts, which, hand in hand with genomic sequencing, will help improve fungal bioinformatics databases for future studies of Botryosphaeriaceae. All data, including raw data, are available via the ProteomeXchange data repository with identifier PXD005283. This is the first study of its kind in Botryosphaeriaceae.
关于病原体中蛋白质功能,尤其是在枝干病害真菌家族葡萄座腔菌科中的蛋白质功能,仍存在许多基础科学问题,该菌科对经济上重要的植物,尤其是果树来说是一个全球性问题。蛋白质组学是研究蛋白质表达以及在未测序和注释不佳的生物体中发现新蛋白质的一项非常有用的技术。目前的真菌蛋白质组学方法涉及二维十二烷基硫酸钠聚丙烯酰胺凝胶电泳(2D SDS - PAGE)以及广泛、复杂的蛋白质提取方法。在这项工作中,一种改良的福尔克提取法被应用于蛋白质提取,以对植物和人类真菌病原体进行肽测序以及肽片段分析/蛋白质鉴定。两种生物信息学方法都从在存在外源甘油三酯和葡萄糖的情况下产生的蛋白质中获得了新的肽序列。由于这些蛋白质及其可能具有的功能在生物技术中的潜在用途,以及作为理解真菌生物化学的新范例,例如发现变应原性烯醇酶以及各种新型蛋白酶,包括与蛇毒中发现的锌金属蛋白酶同源的蛋白酶,它们可作为进一步功能表征和验证工作的目标。这项工作有助于基因组注释工作,该工作与基因组测序携手,将有助于改进葡萄座腔菌科未来研究的真菌生物信息学数据库。所有数据,包括原始数据,可通过蛋白质组交换数据存储库获取,标识符为PXD005283。这是葡萄座腔菌科此类研究中的首例。