Complementary proteome and glycoproteome access revealed through comparative analysis of reversed phase and porous graphitic carbon chromatography.
Complementary proteome and glycoproteome access revealed through comparative analysis of reversed phase and porous graphitic carbon chromatography.
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通过反相色谱和多孔石墨碳色谱的比较分析揭示了互补蛋白质组和糖蛋白质组的通路。
DOI：
10.1007/s00216-022-03934-7
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发表时间：
2022-07
影响因子：
4.3
通讯作者：
Li, Lingjun
中科院分区：
文献类型：
--
作者：
Delafield, Daniel G.;Miles, Hannah N.;Liu, Yuan;Ricke, William A.;Li, Lingjun
仪器和分析技术的不断发展有助于在蛋白质糖基化和人类疾病之间建立严格的联系。这些疾病，如各种形式的癌症，往往与穷人的血糖，促使需要更全面的糖蛋白质组的表征。虽然创新的仪器和计算策略在很大程度上有利于糖蛋白质组学分析，但较少关注通过替代的优化色谱技术获得的益处。多孔石墨碳（PGC）色谱由于其移动的相灵活性、增加极性分析物的保留和改善在较高温度下的结构解析而在糖组学研究中获得了相当大的兴趣。在高通量自下而上的糖蛋白组学实验中，PGC尚未与标准反相液相色谱（RPLC）进行系统性比较或串联，从而未探索潜在的益处。在一系列柱温下对单相和两相分离方案进行比较分析，说明了每种方法的互补优势。PGC分离显示出选择性地保留较短的、更亲水的糖肽种类，赋予更高的平均电荷，并且对于所鉴定的糖位点表现出更大的微观异质性覆盖。此外，我们证明，糖肽异构体的液相分离可以通过单相和两相PGC分离来实现，提供了一种方法，实现简便，多维糖肽表征。除此之外，我们还展示了如何利用多种分离制度和柱温可以帮助分析致瘤性和侵袭性前列腺癌细胞中的糖蛋白组。RAW MS蛋白质组学和糖蛋白质组学数据集已通过PRIDE合作伙伴存储库存放到ProteomeXchange Consortium，数据集标识符分别为PXD 024196（10.6019/PXD 024196）和PXD 024195。
Continual development in instrumental and analytical techniques have aided in establishing rigorous connections between protein glycosylation and human illness. These illnesses, such as various forms of cancer, are often associated with poor prognoses, prompting the need for more comprehensive characterization of the glycoproteome. While innovative instrumental and computational strategies have largely benefited glycoproteomic analyses, less attention is given to benefits gained through alternative, optimized chromatographic techniques. Porous graphitic carbon (PGC) chromatography has gained considerable interest in glycomics research due to its mobile phase flexibility, increased retention of polar analytes and improved structural elucidation at higher temperatures. PGC has yet to be systematically compared against or in tandem with standard reversed phase liquid chromatography (RPLC) in high-throughput bottom-up glycoproteomics experiments, leaving the potential benefits unexplored. Performing comparative analysis of single and biphasic separation regimes at a range of column temperatures illustrates complementary advantages for each method. PGC separation is shown to selectively retain shorter, more hydrophilic glycopeptide species, imparting higher average charge, and exhibiting greater microheterogeneity coverage for identified glycosites. Additionally, we demonstrate that liquid-phase separation of glycopeptide isomers may be achieved through both single and biphasic PGC separations, providing a means towards facile, multidimensional glycopeptide characterization. Beyond this, we demonstrate how utilization of multiple separation regimes and column temperatures can aid in profiling the glycoproteome in tumorigenic and aggressive prostate cancer cells. RAW MS proteomics and glycoproteomics datasets have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD024196 (10.6019/PXD024196) and PXD024195, respectively.
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影响因子：
7.3
作者：
Hobbs SJ;Nolz JC
通讯作者：
Nolz JC
影响因子：
7.8
作者：
Chen Z;Glover MS;Li L
通讯作者：
Li L
影响因子：
4.6
作者：
Losev, Yelena;Paul, Ashim;Segal, Daniel
通讯作者：
Segal, Daniel
DOI：
10.1073/pnas.1916498117
发表时间：
2020-04-07
影响因子：
11.1
作者：
Park, Diane Dayoung;Phoomak, Chatchai;Lebrilla, Carlito B.
通讯作者：
Lebrilla, Carlito B.
DOI：
10.1007/s13361-018-1932-z
发表时间：
2018-06-01
影响因子：
3.2
作者：
Ashwood, Christopher;Lin, Chi-Hung;Packer, Nicolle H.
通讯作者：
Packer, Nicolle H.