Proteomic analysis of the honey bee worker venom gland focusing on the mechanisms of protection against tissue damage
Proteomic analysis of the honey bee worker venom gland focusing on the mechanisms of protection against tissue damage
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DOI：
10.1016/j.toxicon.2008.05.003
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发表时间：
2008-07-01
期刊：
影响因子：
2.8
通讯作者：
Jacobs, Frans J.
中科院分区：
文献类型：
--
作者：
Peiren, Nico;de Graaf, Dirk C.;Jacobs, Frans J.
当蜜蜂面临危险和捕食者时，工蜂会使用毒液来保护蜂群和自己。它由细长、曲折且分叉的腺体产生，由多种有毒蛋白质和肽组成。本研究的目的是确定保护毒腺分泌细胞免受这些有害成分侵害的机制。通过二维凝胶电泳分离整个毒腺样品，包括互连的储库，并使用 MALDI TOF/TOF-MS 和 LC MS/MS 对最丰富的蛋白质点进行质谱鉴定。这项蛋白质组学研究揭示了四种抗氧化酶：CuZn超氧化物歧化酶 (SOD1)、谷胱甘肽-S-转移酶 sigma 1 同种型 A (GSTS1)、过氧化还原蛋白 2540 (PXR2540) 和硫氧还蛋白过氧化物酶 I 同种型 A (TPX1)。尽管谷胱甘肽-S-转移酶 (GST) 也与异生物质解毒有关，但我们发现的蛋白质属于 GST Sigma 类，已知该蛋白仅能防止氧化应激。此外，我们还可以证明，与成年蜜蜂的其他组织相比，毒腺的 GST 和 SOD 活性分别较低和中等。同样发现了几种与其他形式的应激有关的蛋白质，但仍不确定它们在毒腺中的功能。除了先前蛋白质组学研究中已发现的主要蜂王浆蛋白 9 (MRJP9) 之外，我们还确定 MRJP8 是与毒腺相关的 MRJP 蛋白家族的第二个成员。两个 MRJP 的转录本均被扩增并测序。 2D 凝胶中大量存在两种表皮内结构蛋白，很可能代表表皮内层的结构成分，保护分泌细胞免受其产生的毒素的侵害。 (C) 2008 Elsevier Ltd. 保留所有权利。
Honey bee workers use venom for the defence of the colony and themselves when they are exposed to dangers and predators. It is produced by a long thin, convoluted, and bifurcated gland, and consists of several toxic proteins and peptides. The present study was undertaken in order to identify the mechanisms that protect the venom gland secretory cells against these harmful components. Samples of whole venom glands, including the interconnected reservoirs, were separated by two-dimensional gel electrophoresis and the most abundant protein spots were subjected to mass spectrometric identification using MALDI TOF/TOF-MS and LC MS/MS. This proteomic study revealed four antioxidant enzymes: CuZn superoxide dismutase (SOD1), glutathione-S-transferase sigma 1 isoform A (GSTS1), peroxiredoxin 2540 (PXR2540) and thioredoxin peroxidase I isoform A (TPX1). Although glutathione-S-transferase (GST) has also been associated with xenobiotic detoxification, the protein we found belongs to the GST Sigma class which is known to protect against oxidative stress only. Moreover, we could demonstrate that the GST and SOD activity of the venom gland was low and moderate, respectively, when compared to other tissues from the adult honey bee. Several proteins involved in other forms of stress were likewise found but it remains uncertain what their function is in the venom gland. In addition to major royal jelly protein 9 (MRJP9), already found in a previous proteomic study, we identified MRJP8 as second member of the MRJP protein family to be associated with the venom gland. Transcripts of both MRJPs were amplified and sequenced. Two endocuticular structural proteins were abundantly present in the 2D-gel and most probably represent a structural component of the epicuticular lining that protects the secretory cells from the toxins they produce. (C) 2008 Elsevier Ltd. All rights reserved.