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Identification and Quantitation of Amylase Trypsin Inhibitors Across Cultivars Representing the Diversity of Bread Wheat.
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2020-04-08 , DOI: 10.1021/acs.jproteome.0c00059
Utpal Bose 1 , Angéla Juhász 2 , James A Broadbent 1 , Keren Byrne 1 , Crispin A Howitt 3 , Michelle L Colgrave 1, 2
Affiliation  

α-Amylase/trypsin inhibitors (ATIs) may have a role in nonceliac wheat sensitivity (NCWS) and celiac disease (CD), but the ATI content and diversity across a range of wheat cultivars are not well characterized. Discovery proteomics was used to detect ATIs across two wheat cultivars: Chara and Magenta. Comprehensive mapping of detected ATIs with the ATIs from the recently published wheat genome RefSeq v1.0 shows the presence of three major subclasses: monomeric (9%), dimeric (61%), and chloroform-methanol (CM) type (30%). Subsequently, the level of 18 ATI isoforms (63 peptides) grouped into four subtypes was monitored across 15 commercial wheat cultivars and the eight parental lines from a multiparent advanced-generation intercross (MAGIC) population using liquid chromatography-multiple reaction monitoring-mass spectrometry (LC-MRM-MS). The ATI content of wheat cultivars Janz, Sunvale, Diamond Bird, and Longreach Scout was significantly lower than that of other wheat cultivars. The MAGIC parental cultivars Baxter and Xiaoyan 54 contain higher levels (∼115% relative to the average wheat ATI content), whereas cultivar Pastor contained the lowest levels (∼87%). Comprehensive sequence analysis, annotation, chromosomal locations, and epitope mapping enabled us to build an LC-MRM-MS method to monitor and quantify the immunostimulatory ATI proteins potentially related to NCWS, autoimmune diseases, and metabolic disorders. This provides an opportunity to select wheat cultivars with significantly lower levels of ATIs.

中文翻译:

代表面包小麦多样性的品种间淀粉酶胰蛋白酶抑制剂的鉴定和定量。

α-淀粉酶/胰蛋白酶抑制剂 (ATI) 可能在非腹腔小麦敏感性 (NCWS) 和乳糜泻 (CD) 中起作用,但一系列小麦品种的 ATI 含量和多样性尚未得到很好的表征。Discovery 蛋白质组学用于检测两种小麦品种的 ATI:Chara 和 Magenta。检测到的 ATI 与最近发表的小麦基因组 RefSeq v1.0 中的 ATI 的综合作图显示存在三个主要亚类:单体 (9%)、二聚体 (61%) 和氯仿-甲醇 (CM) 型 (30%) . 随后,使用液相色谱-多反应监测-质谱法监测了 15 个商业小麦品种和来自多亲先进代杂交 (MAGIC) 群体的 8 个亲本系的 18 个 ATI 同种型(63 种肽)的水平,这些亚型分为四个亚型。 LC-MRM-MS)。小麦品种Janz、Sunvale、Diamond Bird和Longreach Scout的ATI含量显着低于其他小麦品种。MAGIC 亲本品种 Baxter 和小燕 54 的含量较高(~115% 相对于小麦 ATI 的平均含量),而品种 Pastor 的含量最低(~87%)。全面的序列分析、注释、染色体位置和表位作图使我们能够建立一种 LC-MRM-MS 方法来监测和量化可能与 NCWS、自身免疫性疾病和代谢紊乱相关的免疫刺激性 ATI 蛋白。这为选择 ATI 水平显着降低的小麦品种提供了机会。MAGIC 亲本品种 Baxter 和小燕 54 的含量较高(~115% 相对于小麦 ATI 的平均含量),而品种 Pastor 的含量最低(~87%)。全面的序列分析、注释、染色体位置和表位作图使我们能够建立一种 LC-MRM-MS 方法来监测和量化可能与 NCWS、自身免疫性疾病和代谢紊乱相关的免疫刺激性 ATI 蛋白。这为选择 ATI 水平显着降低的小麦品种提供了机会。MAGIC 亲本品种 Baxter 和小燕 54 的含量较高(~115% 相对于小麦 ATI 的平均含量),而品种 Pastor 的含量最低(~87%)。全面的序列分析、注释、染色体位置和表位作图使我们能够建立一种 LC-MRM-MS 方法来监测和量化可能与 NCWS、自身免疫性疾病和代谢紊乱相关的免疫刺激性 ATI 蛋白。这为选择 ATI 水平显着降低的小麦品种提供了机会。和表位作图使我们能够建立一种 LC-MRM-MS 方法来监测和量化可能与 NCWS、自身免疫性疾病和代谢紊乱相关的免疫刺激性 ATI 蛋白。这为选择 ATI 水平显着降低的小麦品种提供了机会。和表位作图使我们能够建立一种 LC-MRM-MS 方法来监测和量化可能与 NCWS、自身免疫性疾病和代谢紊乱相关的免疫刺激性 ATI 蛋白。这为选择 ATI 水平显着降低的小麦品种提供了机会。
更新日期:2020-04-08
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