《自然—方法學(xué)》介紹了一種提高質(zhì)譜儀分辨率和可用性的改進(jìn)方法,,科研人員可利用該方法獲得完整蛋白復(fù)合物的重要結(jié)構(gòu)信息。
關(guān)于大型蛋白復(fù)合物的詳細(xì)結(jié)構(gòu)信息包括可與之結(jié)合的分子有哪些,、該復(fù)合物是如何組成的等,是可以通過使用能檢測高分子量離子的質(zhì)譜技術(shù)來獲得的,。迄今為止,,這項(xiàng)技術(shù)的實(shí)現(xiàn)需要采用特殊定制的質(zhì)譜儀,且僅有少數(shù)專業(yè)實(shí)驗(yàn)室能辦到,。
Albert Heck等人通過改裝軌道阱質(zhì)譜儀這種被廣泛用于高通量蛋白譜的常用儀器,,成功地實(shí)現(xiàn)了對最高分子量可達(dá)一百萬摩爾質(zhì)量的完整蛋白復(fù)合物的檢測。經(jīng)過他們改裝的質(zhì)譜儀具有足夠高的分辨率,,可識別出個體ATP或ADP分子與細(xì)菌的伴侶蛋白復(fù)合物GroEL的結(jié)合——目前的無損質(zhì)譜技術(shù)還無法做到這一點(diǎn),。該方法同樣能夠應(yīng)用于蛋白復(fù)合物轉(zhuǎn)錄后修飾的靈敏度檢測。(生物谷Bioon.com)
doi:10.1038/nmeth.2208
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High-sensitivity Orbitrap mass analysis of intact macromolecular assemblies
Rebecca J Rose, Eugen Damoc, Eduard Denisov, Alexander Makarov & Albert J R Heck
The analysis of intact protein assemblies in native-like states by mass spectrometry offers a wealth of information on their biochemical and biophysical properties. Here we show that the Orbitrap mass analyzer can be used to measure protein assemblies of molecular weights approaching one megadalton with sensitivity down to the detection of single ions. Minor instrumental modifications enabled the measurement of various protein assemblies with outstanding mass-spectral resolution.