Scanning Quadrupole Data-Independent Acquisition, Part B: Application to the Analysis of the Calcineurin-Interacting Proteins during Treatment of Aspergillus fumigatus with Azole and Echinocandin Antifungal Drugs.
Scanning Quadrupole Data-Independent Acquisition, Part B: Application to the Analysis of the Calcineurin-Interacting Proteins during Treatment of Aspergillus fumigatus with Azole and Echinocandin Antifungal Drugs.
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DOI：
10.1021/acs.jproteome.7b00499
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发表时间：
2018-02-02
影响因子：
4.4
通讯作者：
Steinbach WJ
中科院分区：
文献类型：
--
作者：
Juvvadi PR;Moseley MA;Hughes CJ;Soderblom EJ;Lennon S;Perkins SR;Thompson JW;Geromanos SJ;Wildgoose J;Richardson K;Langridge JI;Vissers JPC;Steinbach WJ
钙调神经磷酸酶是一种重要的细胞信号蛋白，在几种真菌病原体中协调生长、应激反应、毒力和抗真菌药物抗性。阻断钙调神经磷酸酶信号传导增加了几种目前可用的抗真菌药物的疗效，并抑制耐药性。我们展示了一种新的扫描四极DIA方法的应用程序，用于分析致命的人类真菌病原体烟曲霉的治疗性抗真菌药物治疗过程中与钙调磷酸酶免疫共沉淀的蛋白质的变化。我们的实验设计提供了对方法精密度的评估，如通过对研究合并液QC样品的8份重复样品进行以肽和蛋白质为中心的分析所证明的。两种不同类别的临床相关的抗真菌药物，这是指南推荐用于治疗由烟曲霉，唑类（伏立康唑）和棘白菌素类（卡泊芬净和米卡芬净）引起的侵袭性“曲霉病”，它们分别特异性靶向真菌质膜和真菌细胞壁，被选择来区分与钙调磷酸酶共免疫沉淀的蛋白质中发生的变化。新的潜在的相互作用被确定在响应于不同的药物治疗，这是钙调神经磷酸酶在调节这些效应的可能作用的指示。值得注意的是，伏立康唑治疗显示参与膜麦角固醇生物合成的关键蛋白质与钙调磷酸酶的免疫沉淀增加。与此相反，棘白菌素（卡泊芬净或米卡芬净）治疗引起增加参与细胞壁生物合成和分隔的蛋白质的免疫沉淀。此外，丰富的免疫共沉淀的核糖体蛋白与钙调磷酸酶只发生在棘白菌素治疗，表明在不同的抗真菌药物治疗的细胞生长机制的重编程。虽然所观察到的钙调神经磷酸酶免疫沉淀蛋白的变化也可能是由于不同药物治疗下其表达水平的变化，但本研究表明钙调神经磷酸酶依赖性细胞机制在抗真菌治疗A.值得进一步研究的烟曲霉。
Calcineurin is a critical cell-signaling protein that orchestrates growth, stress response, virulence, and antifungal drug resistance in several fungal pathogens. Blocking calcineurin signaling increases the efficacy of several currently available antifungals and suppresses drug resistance. We demonstrate the application of a novel scanning quadrupole DIA method for the analysis of changes in the proteins coimmunoprecipitated with calcineurin during therapeutic antifungal drug treatments of the deadly human fungal pathogen Aspergillus fumigatus. Our experimental design afforded an assessment of the precision of the method as demonstrated by peptide- and protein-centric analysis from eight replicates of the study pool QC samples. Two distinct classes of clinically relevant antifungal drugs that are guideline recommended for the treatment of invasive “aspergillosis” caused by Aspergillus fumigatus, the azoles (voriconazole) and the echinocandins (caspofungin and micafungin), which specifically target the fungal plasma membrane and the fungal cell wall, respectively, were chosen to distinguish variations occurring in the proteins coimmunoprecipitated with calcineurin. Novel potential interactors were identified in response to the different drug treatments that are indicative of the possible role for calcineurin in regulating these effectors. Notably, treatment with voriconazole showed increased immunoprecipitation of key proteins involved in membrane ergosterol biosynthesis with calcineurin. In contrast, echinocandin (caspofungin or micafungin) treatments caused increased immunoprecipitation of proteins involved in cell-wall biosynthesis and septation. Furthermore, abundant coimmunoprecipitation of ribosomal proteins with calcineurin occurred exclusively in echinocandins treatment, indicating reprogramming of cellular growth mechanisms during different antifungal drug treatments. While variations in the observed calcineurin immunoprecipitated proteins may also be due to changes in their expression levels under different drug treatments, this study suggests an important role for calcineurin-dependent cellular mechanisms in response to antifungal treatment of A. fumigatus that warrants future studies.
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影响因子：
4.9
作者：
Juvvadi PR;Muñoz A;Lamoth F;Soderblom EJ;Moseley MA;Read ND;Steinbach WJ
通讯作者：
Steinbach WJ
影响因子：
4.9
作者：
Cruz, MC;Del Poeta, M;Heitman, J
通讯作者：
Heitman, J
影响因子：
5.8
作者：
Dinamarco, Taisa Magnani;Brown, Neil Andrew;Goldman, Gustavo Henrique
通讯作者：
Goldman, Gustavo Henrique
影响因子：
2.8
作者：
Jiang, Hechun;Ouyang, Haomiao;Jin, Cheng
通讯作者：
Jin, Cheng
影响因子：
4.9
作者：
Gautam, Poonam;Shankar, Jata;Sarma, Puranam Usha
通讯作者：
Sarma, Puranam Usha