Absolute Quantitation of Tryptophan-Containing Peptides and Amyloid β-Peptide Fragments by Coulometric Mass Spectrometry.
Absolute Quantitation of Tryptophan-Containing Peptides and Amyloid β-Peptide Fragments by Coulometric Mass Spectrometry.
复制标题
库仑质谱法测定含色氨酸肽和淀粉样β肽片段的绝对定量。
DOI：
10.1021/jasms.1c00121
复制
发表时间：
2021-07-07
影响因子：
3.2
通讯作者：
Chen H
中科院分区：
文献类型：
--
作者：
Ai Y;Zhao P;Fnu PIJ;Chen H
同位素标记的内标通常用于基于质谱仪(MS)的绝对定量。然而，合成同位素标记的多肽是耗时和昂贵的。为了解决这个问题，我们最近开发了一种库仑质谱法(CMS)，用于在不使用标准的情况下进行绝对定量，该方法基于半胱氨酸或酪氨酸多肽的电化学氧化，然后用质谱仪测量氧化产率。为了进一步扩大该方法的应用范围，本文提出了基于色氨酸电化学氧化的CMS方法用于多肽的绝对定量。WGG、WQPPRARI、WAGGDASGE、RTRPLWVRME和KVPRNQDWL等色氨酸多肽的定量误差为−4.5%~+4.3%。此外，这种定量方法也适用于蛋白质，即蛋白质可以被消化，可以选择一个替代多肽来定量反映母体蛋白质的含量，以细胞色素c为原料的多肽GITWK的CMS分析为例，CMS结果与传统的同位素稀释法吻合良好，仅有3.5%的微小差异。此外，还利用CMS成功地定量了基于酪氨酸氧化的淀粉样β蛋白(Aβ)多肽片段(最多28个氨基酸残基)。CMS方法在不使用同位素标记的多肽标准品的情况下对多肽和蛋白质绝对定量的有效性将极大地促进蛋白质组学的研究。
Isotope labeled internal standards are routinely used for mass spectrometry (MS)-based absolute quantitation. However, syntheses of isotope-labeled peptides are time-consuming and costly. To tackle this issue, we recently developed a coulometric mass spectrometric (CMS) approach for absolute quantitation without the use of standards, based on the electrochemical oxidation of cysteine or tyrosine-containing peptides followed with mass spectrometric measurement of the oxidation yield. To further expand the utility of this method, herein, we present the CMS method for absolute quantitation of peptides based on tryptophan electrochemical oxidation. Several tryptophan-containing peptides, such as WGG, WQPPRARI, WAGGDASGE, RTRPLWVRME, and KVPRNQDWL were successfully quantified with a quantification error ranging from −4.5 to +4.3%. Furthermore, this quantitation approach is also applicable to protein, in which protein can be digested and a surrogate peptide can be selected for quantification to reflect the amount of the parent protein, as exemplified by CMS analysis of peptide GITWK from cytochrome c. The CMS result agreed well with the traditional isotope dilution method, with only a small difference of 3.5%. In addition, CMS was used to successfully quantify amyloid beta (Aβ) peptide fragments (up to 28 amino acid residues) based on tyrosine oxidation. The validity of CMS method for peptide and protein absolute quantitation without using isotope-labeled peptide standards would greatly facilitate proteomics research.
登录
查看更多内容
影响因子：
2.9
作者：
Lame, Mary E.;Chambers, Erin E.;Blatnik, Matthew
通讯作者：
Blatnik, Matthew
影响因子：
9.9
作者：
Lange, Vinzenz;Picotti, Paola;Domon, Bruno;Aebersold, Ruedi
通讯作者：
Aebersold, Ruedi
影响因子：
168.9
作者：
Ferri, CP;Prince, M;Scazufca, M
通讯作者：
Scazufca, M
常染色体主导的阿尔茨海默氏病中CSF生物标志物的纵向变化。
影响因子：
17.1
作者：
Fagan AM;Xiong C;Jasielec MS;Bateman RJ;Goate AM;Benzinger TL;Ghetti B;Martins RN;Masters CL;Mayeux R;Ringman JM;Rossor MN;Salloway S;Schofield PR;Sperling RA;Marcus D;Cairns NJ;Buckles VD;Ladenson JH;Morris JC;Holtzman DM;Dominantly Inherited Alzheimer Network
通讯作者：
Dominantly Inherited Alzheimer Network
影响因子：
7.4
作者：
Roeser, Julien;Permentier, Hjalmar P.;Bischoff, Rainer
通讯作者：
Bischoff, Rainer