First protein and peptide characterization of the tarsal adhesive secretions in the desert locust, Schistocerca gregaria, and the Madagascar hissing cockroach, Gromphadorhina portentosa
First protein and peptide characterization of the tarsal adhesive secretions in the desert locust, Schistocerca gregaria, and the Madagascar hissing cockroach, Gromphadorhina portentosa
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DOI：
10.1111/imb.12241
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发表时间：
2016-10-01
影响因子：
2.6
通讯作者：
Neuenfeldt, M.
中科院分区：
文献类型：
--
作者：
Betz, O.;Maurer, A.;Neuenfeldt, M.
在昆虫跗骨黏附物的分析中，多肽和蛋白质在很大程度上被忽略了。在提取沙漠蝗（Schistocerca gregaria）和马达加斯加嘶叫蠊（Gromphadorhina portentosa）跗分泌物蛋白组分的基础上，结合傅里叶变换红外光谱（FTIR）、十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）和基质辅助激光解吸/电离质谱（MALDI-TOF MS）技术进行蛋白质量检测。在这两种昆虫中，SDS-PAGE分析显示，跗骨分泌物和胫骨对照样品中都有8-190 kDa的蛋白带。两个（S. gregaria）和一个（G. portentosa）蛋白带只出现在跗骨分泌物中，可以认为属于这种分泌物所特有的肽和蛋白质。MALDITOF分析显示，在S. gregaria中有83种1-7 kDa的不同蛋白/肽，在G. portentosa中有48种1-11 kDa的不同蛋白/肽。59个（S. gregaria）和27个（G. portentosa）蛋白只存在于跗骨分泌物中。在G. portentosa中，在c. 10-12 kDa范围内出现了一系列特征信号峰，每个峰相距约160 Da。这种模式表明蛋白质经过糖基化修饰。我们的方法表明，广泛的采样涉及大量的时间和人力，直接从跗骨中取样黏附液，为提取足够数量的肽和蛋白质开辟了一个前景。这使得它们可以进入蛋白质组学领域，从而阐明它们在粘附过程中的可能功能。
Peptides and proteins have been largely neglected in the analysis of insect tarsal adhesives. After extraction of the protein fraction of the tarsal secretion of the desert locust, Schistocerca gregaria, and Madagascar hissing cockroach, Gromphadorhina portentosa, we combined Fourier transform infrared spectroscopy (FTIR), sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) analyses for protein mass detection. In both these insects, SDS-PAGE analysis revealed several protein bands ranging from 8-190 kDa in both the tarsal secretion and the tibia control sample. Two (S. gregaria) and one (G. portentosa) protein bands exclusively occurred in the tarsal secretion and can be considered to belong to peptides and proteins specific to this secretion. MALDITOF analyses revealed 83 different proteins/peptides of 1-7 kDa in S. gregaria, and 48 of 1-11 kDa in G. portentosa. 59 (S. gregaria) and 27 (G. portentosa) proteins exclusively occurred in the tarsal secretion. In G. portentosa, a characteristic series of signal peaks occurred in the range of c. 10-12 kDa, each peak being approximately 160 Da apart. Such a pattern is indicative of proteins modified by glycosylation. Our approach demonstrates that extensive sampling involving considerable time and manpower to sample the adhesive fluid directly from the tarsi opens up a perspective for extracting peptides and proteins in sufficient quantities. This makes them accessible to the field of proteomics and thus to elucidate their possible function in the adhesive process.