New Enzymatic Approach to Distinguish Fucosylation Isomers of N-Linked Glycans in Tissues Using MALDI Imaging Mass Spectrometry.
New Enzymatic Approach to Distinguish Fucosylation Isomers of N-Linked Glycans in Tissues Using MALDI Imaging Mass Spectrometry.
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DOI：
10.1021/acs.jproteome.0c00024
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发表时间：
2020-08-07
影响因子：
4.4
通讯作者：
Mehta AS
中科院分区：
文献类型：
--
作者：
West CA;Liang H;Drake RR;Mehta AS
N-连接聚糖的特定改变，如核心岩藻糖基化，与许多癌症和其他疾病状态相关。由于与岩藻糖基化N-聚糖相关的许多可能的异头键，确定特定的异头键和岩藻糖基化位点（即，芯相对于外臂）可能难以阐明。使用重组内切糖苷酶F3（Endo F3）描述了福尔马林固定的临床组织中新的MALDI质谱成像工作流程，该酶具有切割附着于糖蛋白的核心岩藻糖基化N-聚糖的特定偏好。与更广泛的底物酶肽-N-糖苷酶F（PNGaseF）相比，Endo F3在蛋白质附着位点的两个核心N-乙酰葡糖胺残基之间切割。在组织上，这导致349.137 a. m. u的质量偏移。与标准PNGaseF释放的N-聚糖相比，核心岩藻糖基化的N-聚糖。Endo F3可以单独使用，也可以与PNGaseF消化相同的组织切片组合使用。肝脏和前列腺组织中的初步结果表明，核心岩藻糖基化聚糖与特定组织区域相关，同时仍证明在所有组织区域中存在核心和外臂岩藻糖基化聚糖的多样化混合。通过确定这些特定的连接，同时保持定位，更有针对性的疾病状态的诊断生物标志物是可能的，而不需要显微解剖或溶解的组织。
Specific alterations in N-linked glycans, such as core fucosylation, are associated with many cancers and other disease states. Because of the many possible anomeric linkages associated with fucosylated N-glycans, determination of specific anomeric linkages and the site of fucosylation (i.e., core vs outer arm) can be difficult to elucidate. A new MALDI mass spectrometry imaging workflow in formalin-fixed clinical tissues is described using recombinant endoglycosidase F3 (Endo F3), an enzyme with a specific preference for cleaving core-fucosylated N-glycans attached to glycoproteins. In contrast to the broader substrate enzyme peptide-N-glycosidase F (PNGaseF), Endo F3 cleaves between the two core N-acetylglucosamine residues at the protein attachment site. On tissues, this results in a mass shift of 349.137 a.m.u. for core-fucosylated N-glycans when compared to N-glycans released with standard PNGaseF. Endo F3 can be used singly and in combination with PNGaseF digestion of the same tissue sections. Initial results in liver and prostate tissues indicate core-fucosylated glycans associated to specific tissue regions while still demonstrating a diverse mix of core- and outer arm-fucosylated glycans throughout all regions of tissue. By determining these specific linkages while preserving localization, more targeted diagnostic biomarkers for disease states are possible without the need for microdissection or solubilization of the tissue.
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DOI：
10.1158/1078-0432.ccr-07-5261
发表时间：
2009-03-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Goldman R;Ressom HW;Varghese RS;Goldman L;Bascug G;Loffredo CA;Abdel-Hamid M;Gouda I;Ezzat S;Kyselova Z;Mechref Y;Novotny MV
通讯作者：
Novotny MV
DOI：
10.1073/pnas.79.15.4540
发表时间：
1982-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
ELDER, JH;ALEXANDER, S
通讯作者：
ALEXANDER, S
影响因子：
7
作者：
Kirmiz, Crystal;Li, Bensheng;Miyamoto, Suzanne
通讯作者：
Miyamoto, Suzanne
影响因子：
8
作者：
Andrianifahanana, M;Agrawal, A;Batra, SK
通讯作者：
Batra, SK
影响因子：
7
作者：
Karamessinis, Panagiotis M.;Malamitsi-Puchner, Ariadne;Chrousos, George
通讯作者：
Chrousos, George