Site-Specific Intact N-Linked Glycopeptide Characterization of Prostate-Specific Membrane Antigen from Metastatic Prostate Cancer Cells.
Site-Specific Intact N-Linked Glycopeptide Characterization of Prostate-Specific Membrane Antigen from Metastatic Prostate Cancer Cells.
复制标题
DOI：
10.1021/acsomega.2c02265
复制
发表时间：
2022-08-30
期刊：
影响因子：
4.1
通讯作者：
Nyalwidhe, Julius O.
中科院分区：
文献类型：
--
作者：
Mackay, Stephen;Hitefield, Naomi L.;Oduor, Ian O.;Roberts, Autumn B.;Burch, Tanya C.;Lance, Raymond S.;Cunningham, Tina D.;Troyer, Dean A.;Semmes, Oliver J.;Nyalwidhe, Julius O.
与前列腺特异性膜抗原(PSMA)偶联的N-连接糖链的组成及其在前列腺癌进展中的功能意义尚未完全确定。PSMA是从两个具有不同组织趋向性和定位的转移性前列腺癌细胞系LNCaP和MDAPCa2b中分离得到的。分离的PSMA经胰酶消化，在三元Orbitrap Fusion Lumos质谱仪上进行LC-HCD-EThcD-MS/MS分析。用Byonic和Byology软件对位点特异性N-糖肽进行定性和定量的差异性分析。比较定量分析表明，天冬酰胺残基51、76、121、195、336、459、476和638处的多个糖肽在两个细胞系中的丰度存在显著差异(p<；0.05)。生化分析使用内切糖苷酶处理和凝集素捕获确认MS和站点占有率数据。这些数据证明了PSMA糖肽综合分析策略的有效性。这一方法将成为正在进行的实验的基础，以确定从疾病分层的临床样本中分离的PSMA中特定部位的糖链变化，以揭示可能与疾病进展和转移表型相关的靶点。
The composition of N-linked glycans that are conjugated to the prostate-specific membrane antigen (PSMA) and their functional significance in prostate cancer progression have not been fully characterized. PSMA was isolated from two metastatic prostate cancer cell lines, LNCaP and MDAPCa2b, which have different tissue tropism and localization. Isolated PSMA was trypsin-digested, and intact glycopeptides were subjected to LC-HCD-EThcD-MS/MS analysis on a Tribrid Orbitrap Fusion Lumos mass spectrometer. Differential qualitative and quantitative analysis of site-specific N-glycopeptides was performed using Byonic and Byologic software. Comparative quantitative analysis demonstrates that multiple glycopeptides at asparagine residues 51, 76, 121, 195, 336, 459, 476, and 638 were in significantly different abundance in the two cell lines (p < 0.05). Biochemical analysis using endoglycosidase treatment and lectin capture confirm the MS and site occupancy data. The data demonstrate the effectiveness of the strategy for comprehensive analysis of PSMA glycopeptides. This approach will form the basis of ongoing experiments to identify site-specific glycan changes in PSMA isolated from disease-stratified clinical samples to uncover targets that may be associated with disease progression and metastatic phenotypes.
登录
查看更多内容
影响因子：
7.4
作者：
Toghi Eshghi S;Shah P;Yang W;Li X;Zhang H
通讯作者：
Zhang H
癌症生物标志物发现：靶向糖蛋白的基于凝集素的策略。
影响因子：
--
作者：
Clark D;Mao L
通讯作者：
Mao L
DOI：
10.1002/pros.24254
发表时间：
2022-01
期刊：
The Prostate
影响因子：
--
作者：
通讯作者：
--
影响因子：
2.8
作者：
Ghosh, A;Heston, WDW
通讯作者：
Heston, WDW
影响因子：
4
作者：
Ghosh, A;Heston, WDW
通讯作者：
Heston, WDW