The feeding tube of cyst nematodes: characterisation of protein exclusion.
The feeding tube of cyst nematodes: characterisation of protein exclusion.
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DOI：
10.1371/journal.pone.0087289
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Urwin PE
中科院分区：
文献类型：
--
作者：
Eves-van den Akker S;Lilley CJ;Ault JR;Ashcroft AE;Jones JT;Urwin PE
植物寄生线虫包括几类;其中最具经济破坏性的是定居体内寄生虫。定居的内寄生线虫是专性生物营养体，并使用一套效应蛋白来修饰宿主根组织，以创建作为其唯一营养来源的进食位点。它们通过一种被称为饲管的结构从进食部位吸取宿主细胞的同化物来进食。饲管的功能、组成和分子特征的表征较差。据推测，饲管通过充当分子筛来促进宿主细胞同化物的摄取。几项研究，使用分子量作为蛋白质大小的唯一指标，给出了矛盾的结果，有关的排斥限制的孢囊线虫摄食管。在这项研究中，我们提出了一种方法来预测蛋白质的大小，基于蛋白质数据库的坐标在硅片上。我们使用行波离子迁移谱-质谱法测试了这些预测的有效性，其中预测值和测量值在约6%以内。我们使用的预测，再加上质谱，分析超离心和蛋白质电泳，以解决以前的冲突，并定义的排斥特征的孢囊线虫摄食管。在液相、固相和气相中测试异源性，以提供对饲管尺寸排阻特别感兴趣的三种蛋白质GFP、mRFP和Dual PI的综合评价。本文所述的数据和程序可应用于设计植物表达的防御化合物，旨在吸收到胞囊线虫。我们还强调需要评估蛋白质的异质性时，创造新的融合蛋白。
Plant parasitic nematodes comprise several groups; the most economically damaging of these are the sedentary endoparasites. Sedentary endoparasitic nematodes are obligate biotrophs and modify host root tissue, using a suite of effector proteins, to create a feeding site that is their sole source of nutrition. They feed by withdrawing host cell assimilate from the feeding site though a structure known as the feeding tube. The function, composition and molecular characteristics of feeding tubes are poorly characterised. It is hypothesised that the feeding tube facilitates uptake of host cell assimilate by acting as a molecular sieve. Several studies, using molecular mass as the sole indicator of protein size, have given contradictory results about the exclusion limits of the cyst nematode feeding tube. In this study we propose a method to predict protein size, based on protein database coordinates in silico. We tested the validity of these predictions using travelling wave ion mobility spectrometry – mass spectrometry, where predictions and measured values were within approximately 6%. We used the predictions, coupled with mass spectrometry, analytical ultracentrifugation and protein electrophoresis, to resolve previous conflicts and define the exclusion characteristics of the cyst nematode feeding tube. Heterogeneity was tested in the liquid, solid and gas phase to provide a comprehensive evaluation of three proteins of particular interest to feeding tube size exclusion, GFP, mRFP and Dual PI. The data and procedures described here could be applied to the design of plant expressed defence compounds intended for uptake into cyst nematodes. We also highlight the need to assess protein heterogeneity when creating novel fusion proteins.
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影响因子：
4.4
作者：
Shvartsburg, AA;Schatz, GC;Jarrold, MF
通讯作者：
Jarrold, MF
影响因子：
15
作者：
Mack, E
通讯作者：
Mack, E
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15.9
作者：
GUASCH, A;DEEN, WM;MYERS, BD
通讯作者：
MYERS, BD
影响因子：
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作者：
Mesleh, MF;Hunter, JM;Jarrold, MF
通讯作者：
Jarrold, MF
影响因子：
2.4
作者：
BOCKENHOFF, A;GRUNDLER, FMW
通讯作者：
GRUNDLER, FMW