尽管大多数螨过敏原已在该物种中描述，但屋尘螨Dermatophagoides farinae的过敏原库仍不完整。我们使用蛋白质组学，旨在比较性别之间的蛋白质和过敏原，并为鉴定新型过敏原提供基础。总体上，鉴定出6297个蛋白质命中，分别是男性和女性特异的2899和886。删除跟踪结果会将数据集的范围缩小到3478个匹配项，分别包括275个和157个针对男性和女性的匹配项。所有34个经WHO / IUIS批准的D. farinae鉴定了过敏原（省略了Der f 17），我们还鉴定了尚未描述的Der f 9和38的同源物。Derf 27 / serpin表现出最大的性别依赖性差异，并且在女性中占主导地位。使用官方蛋白质序列，鉴定出Der f 11、14、23、28和30的成功率较低。但是，通过使用较长/完整的序列，Der f 11和14的鉴定成功率大大提高。Der f 30的特征是与此处鉴定的更丰富的Der f 30（同工型）相同的胰蛋白酶消化物。Der f 23在螨体内的丰度似乎很低。分别以低丰度和痕量检测到Der f 28.0101和Der f 28.0201。
意义
在这项工作中，我们对屋尘螨Dermatophagoides farinae进行了蛋白质组学注释，Dermatophagoides farinae是屋尘过敏原的最重要来源。此处进行的蛋白质组学分析不仅为了解螨的生物学特性，而且为鉴定新的过敏原提供了基础。这项研究为D. farinae生成了一个强大的蛋白质组学数据集并审查了该物种中现有的和候选的过敏原。我们强调高通量分析的一些陷阱，尤其是数据库中提供的过敏原蛋白记录的不正确标题可能导致混淆。在蛋白质组学鉴定和定量中使用部分序列会导致鉴定成功率低（信号强度或MS / MS计数低）。因此，我们单独策划了蛋白质序列，以进行正确的鉴定和定量。发现的性别差异可能是影响螨提取物中过敏原/免疫原变异的因素之一。总体而言，这项工作为使用蛋白质组学准确鉴定螨免疫原性蛋白质提供了基准。
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Proteogenomics of the house dust mite, Dermatophagoides farinae: Allergen repertoire, accurate allergen identification, isoforms, and sex-biased proteome differences.
The allergen repertoire of the house dust mite, Dermatophagoides farinae, is incomplete despite most mite allergens having been described in this species. Using proteogenomics, we aimed to compare proteins and allergens between sexes and provide a foundation for the identification of novel allergens. Overall, 6297 protein hits were identified, and 2899 and 886 were male- and female-specific, respectively. Removal of trace results narrowed the dataset to 3478 hits, including 275 and 157 male- and female-specific hits, respectively. All 34 WHO/IUIS-approved D. farinae allergens (omitting Der f 17) were identified, and we also identified homologs of the yet undescribed Der f 9 and 38. Der f 27/serpin exhibited the largest sex-dependent difference and was dominant in females. Using official protein sequences, Der f 11, 14, 23, 28 and 30 were identified with low success. However, identification success of Der f 11 and 14 was greatly increased by using longer/complete sequences. Der f 30 is characterized by the same tryptic digests as the more abundant Der f 30 (isoform) identified here. Der f 23 appears to be of low abundance in mite bodies. Der f 28.0101 and Der f 28.0201 were detected at low abundance and in trace amounts, respectively.
Significance
In this work, we performed a proteogenomic annotation of the house dust mite, Dermatophagoides farinae, which is the most important source of house dust allergens. The proteogenomic analysis performed here provides a foundation for not only understanding the biology of the mite but also the identification of novel allergens. This study generated a robust proteomic dataset for D. farinae and reviewed existing and candidate allergens in this species. We stress some pitfalls of high-throughput analyses, especially that improper headers of allergen protein records provided in databases can lead to confusion. Using partial sequences in proteomic identification and quantification can lead to low identification success (low signal intensity or MS/MS counts). Thus, we individually curated the protein sequences for proper identification and quantification. The discovered sex differences can be one factor affecting allergen/immunogen variations in mite extracts. Overall, this work provides a benchmark for accurate identification of mite immunogenic proteins using proteomics.
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