Quantification of Bioorthogonal Stability in Immune Phagocytes Using Flow Cytometry Reveals Rapid Degradation of Strained Alkynes.
Quantification of Bioorthogonal Stability in Immune Phagocytes Using Flow Cytometry Reveals Rapid Degradation of Strained Alkynes.
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DOI：
10.1021/acschembio.8b00355
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发表时间：
2018-05-18
影响因子：
4
通讯作者：
van Kasteren SI
中科院分区：
文献类型：
--
作者：
Bakkum T;van Leeuwen T;Sarris AJC;van Elsland DM;Poulcharidis D;Overkleeft HS;van Kasteren SI
生物正交化学--在细胞或生物体内进行的化学--已经变得至关重要的领域之一是在研究宿主-病原体相互作用方面。在细胞内病原体的细胞壁或蛋白质组中加入生物正交基，可以在内溶酶体系统内进行研究。然而，为了这种方法的成功，结合的生物正交基必须对这些细胞器中发现的化学条件稳定，这些细胞器是后生动物中最苛刻的：这些基团暴露在氧化物种、酸性条件和活性硫醇中。在这里，我们提出了一种测试，允许评估宿主细胞吞噬体内生物正交群的稳定性。使用基于流式细胞术的分析，我们已经量化了紧张和非紧张的乙炔的树突状细胞吞噬体内的相对标记稳定性。我们发现，在其他系统中表现稳定的基团在吞噬小体成熟后降解了79%。
One of the areas in which bioorthogonal chemistry—chemistry performed inside a cell or organism—has become of pivotal importance is in the study of host–pathogen interactions. The incorporation of bioorthogonal groups into the cell wall or proteome of intracellular pathogens has allowed study within the endolysosomal system. However, for the approach to be successful, the incorporated bioorthogonal groups must be stable to chemical conditions found within these organelles, which are some of the harshest found in metazoans: the groups are exposed to oxidizing species, acidic conditions, and reactive thiols. Here we present an assay that allows the assessment of the stability of bioorthogonal groups within host cell phagosomes. Using a flow cytometry-based assay, we have quantified the relative label stability inside dendritic cell phagosomes of strained and unstrained alkynes. We show that groups that were shown to be stable in other systems were degraded by as much as 79% after maturation of the phagosome.
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影响因子：
15
作者：
Andersen, Kristen A.;Aronoff, Matthew R.;McGrath, Nicholas A.;Raines, Ronald T.
通讯作者：
Raines, Ronald T.
影响因子：
4.7
作者：
Hong, Vu;Steinmetz, Nicole F.;Manchester, Marianne;Finn, M. G.
通讯作者：
Finn, M. G.
影响因子：
18.3
作者：
Hang, Howard C.;Wilson, John P.;Charron, Guillaume
通讯作者：
Charron, Guillaume
DOI：
10.1046/j.1432-1327.2000.01491.x
发表时间：
2000-10-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Furtmüller, PC;Obinger, C;Dunford, HB
通讯作者：
Dunford, HB
影响因子：
15
作者：
Blackman, Melissa L.;Royzen, Maksim;Fox, Joseph M.
通讯作者：
Fox, Joseph M.