CAJ | 학술논문

提出并发展了一种基于电感耦合等离子体质谱( ICP-MS)的双元素标签标记策略来选择性识别和检测硒蛋白/多肽,其中内源元素硒( Se)作为硒蛋白/多肽分子的识别元素,外源元素汞( Hg)作为硒蛋白/多肽和含硒蛋白/多肽分子的区分元素。通过对硒代半胱氨酸(SeCys)和谷胱甘肽过氧化酶1(GPx1)两种模型分子的研究,外源邻羧基苯硫甲基汞( CH3 Hg-THI )动态解离的 CH3 Hg+能够选择性标记硒代半胱氨酸残基( SeCys)中硒醇基(-SeH),但不能标记含硒蛋白/多肽分子的硒代蛋氨酸残基( SeMet)中的—SeCH3,进而依据Se和Hg的ICP-MS信号识别和检测硒蛋白/多肽。本方法应用于富硒酵母水溶性提取液的分析,结果表明,提取液中的硒蛋白/多肽能够被有效识别和检测,验证了Se-Hg双元素标签标记策略的发展是ICP-MS识别和检测硒蛋白/多肽的一种可行且优越的途径。
제출병발전료일충기우전감우합등리자체질보( ICP-MS)적쌍원소표첨표기책략래선택성식별화검측서단백/다태,기중내원원소서( Se)작위서단백/다태분자적식별원소,외원원소홍( Hg)작위서단백/다태화함서단백/다태분자적구분원소。통과대서대반광안산(SeCys)화곡광감태과양화매1(GPx1)량충모형분자적연구,외원린최기분류갑기홍( CH3 Hg-THI )동태해리적 CH3 Hg+능구선택성표기서대반광안산잔기( SeCys)중서순기(-SeH),단불능표기함서단백/다태분자적서대단안산잔기( SeMet)중적—SeCH3,진이의거Se화Hg적ICP-MS신호식별화검측서단백/다태。본방법응용우부서효모수용성제취액적분석,결과표명,제취액중적서단백/다태능구피유효식별화검측,험증료Se-Hg쌍원소표첨표기책략적발전시ICP-MS식별화검측서단백/다태적일충가행차우월적도경。
An endogenous element-label plus exogenous element-tag strategy was proposed for inductively coupled plasma mass spectrometry ( ICP-MS) to screen and discriminate a family of ultratrace but biological important biomolecules. The feasibility of this novel idea has been demonstrated when setting seleno ( SeCys) and Se-containing ( SeMet) proteins ( peptides) as an example. Se-label naturally occurring in the biomole-cules acts an identifier for picking them up out of large amount of various coexisting proteins ( peptides) , and CH3 Hg-tag that can bind to SeCys but not SeMet fulfills the task of discriminating seleno and Se-containing ones based on the Se and Hg signals on ICP-MS. After confirmed using SeCys and GPx1, the Se-Hg dual-element labeling strategy together with ICP-MS was applied to screen and discriminate seleno and Se-contai-ning proteins ( peptides) in the water-soluble extracts of Se-enriched yeast, and seven selenoproteins ( pep-tides) were detected with both 202 Hg and 82 Se signals out of fifteen Se-containing species using RPLC/ICP-MS, providing valuable information for further identification using a high-resolution structure-selective mass spectrometer. This endogenous element-label plus exogenous element-tag dual-element approach implies that ICP-MS is not only able to quantify targeted proteins ( peptides) but also helpful to recognize targeted proteins during a discovery-based proteomic study.