One-step enzymatic modification of the cell surface redirects cellular cytotoxicity and parasite tropism.
One-step enzymatic modification of the cell surface redirects cellular cytotoxicity and parasite tropism.
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DOI：
10.1021/cb500462t
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发表时间：
2015-02-20
影响因子：
4
通讯作者：
Ploegh, Hidde L.
中科院分区：
文献类型：
--
作者：
Swee, Lee Kim;Lourido, Sebastian;Bell, George W.;Ingram, Jessica R.;Ploegh, Hidde L.
工程蛋白的表面展示具有许多有用的应用。由连接至工程化T细胞中的胞质活化基序的细胞外肿瘤特异性抗体片段组成的合成嵌合抗原受体的表达现在被认为是治疗白血病的可行方法。遗传工程细胞转移中固有的从头肿瘤发展的风险要求淋巴细胞质膜功能化的替代方法。我们证明了使用分选酶A在一个步骤中将LPXTG标记的探针和携带LPXTG的蛋白质与质膜上的内源性受体缀合。我们成功地结合生物素探针不仅小鼠造血细胞，但也酵母细胞，293 T细胞，弓形虫。在活化的CD8 T细胞上安装单结构域抗体将细胞特异性细胞毒性重定向至携带相关抗原的细胞。同样地，弓形虫与单结构域抗体的缀合将病原体靶向表达这些单结构域抗体识别的抗原的细胞。这种简单而稳健的酶促方法使得能够在生理反应条件下对质膜进行工程化以用于研究或治疗，从而确保修饰细胞的活力。
Surface display of engineered proteins has many useful applications. The expression of a synthetic chimeric antigen receptor composed of an extracellular tumor-specific antibody fragment linked to a cytosolic activating motif in engineered T cells is now considered a viable approach for the treatment of leukemias. The risk of de novo tumor development, inherent in the transfer of genetically engineered cells, calls for alternative approaches for the functionalization of the lymphocyte plasma membrane. We demonstrate the conjugation of LPXTG-tagged probes and LPXTG-bearing proteins to endogenous acceptors at the plasma membrane in a single step using sortase A. We successfully conjugated biotin probes not only to mouse hematopoietic cells but also to yeast cells, 293T cells, and Toxoplasma gondii. Installation of single domain antibodies on activated CD8 T cell redirects cell-specific cytotoxicity to cells that bear the relevant antigen. Likewise, conjugation of Toxoplasma gondii with single domain antibodies targets the pathogen to cells that express the antigen recognized by these single domain antibodies. This simple and robust enzymatic approach enables engineering of the plasma membrane for research or therapy under physiological reaction conditions that ensure the viability of the modified cells.
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影响因子：
20.3
作者：
Massa, Paul E.;Paniccia, Aida;Rescigno, Maria
通讯作者：
Rescigno, Maria
影响因子：
14.8
作者：
通讯作者：
--
影响因子：
3.3
作者：
Håkansson, S;Morisaki, H;Sibley, LD
通讯作者：
Sibley, LD
DOI：
10.1073/pnas.1101046108
发表时间：
2011-07-12
影响因子：
11.1
作者：
Chen, Irwin;Dorr, Brent M.;Liu, David R.
通讯作者：
Liu, David R.
影响因子：
56.9
作者：
Mahal, LK;Yarema, KJ;Bertozzi, CR
通讯作者：
Bertozzi, CR