Single-molecule and super-resolved imaging deciphers membrane behavior of onco-immunogenic CCR5.
Single-molecule and super-resolved imaging deciphers membrane behavior of onco-immunogenic CCR5.
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单分子和超级分解成像解剖膜的膜行为。
DOI：
10.1016/j.isci.2022.105675
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发表时间：
2022-12-22
期刊：
影响因子：
5.8
通讯作者：
Leake, Mark C.
中科院分区：
文献类型：
--
作者：
Hunter, Patrick;Payne-Dwyer, Alex L.;Shaw, Michael;Signoret, Nathalie;Leake, Mark C.
肿瘤建立致瘤微环境的能力是寻找新疗法的一个重要研究点。肿瘤形成微环境的部分原因是通过趋化轴（如CCR5-CCL5）吸引的免疫细胞的“教育”。此外，癌细胞上调CCR5，再加上其与耐药和转移等致瘤性特征的关联，表明CCR5是一种治疗靶点。然而，随着几个构象“池”被报道，表型研究必须能够揭示构象异质性。为了解决这一挑战，我们在固定细胞中对CCR5进行了超分辨率结构照明显微镜（SIM）和单分子部分tirf偶联HILO （PaTCH）显微镜。SIM数据显示了CCR5的非随机空间分布，而PaTCH图像的CCR5序列的强度跟踪显示了不受CCL5扰动影响的二聚体亚基。这些生物物理方法可以为肿瘤免疫原性受体和许多其他生物分子的结构和功能提供重要的见解。SIM和PaTCH用于CCR5的精确生物成像和单分子跟踪超分辨SIM显示CCR5的非随机聚类单分子PaTCH显示CCR5组装的分子化学计量学；生物物理学;癌症
The ability of tumors to establish a pro-tumorigenic microenvironment is an important point of investigation in the search for new therapeutics. Tumors form microenvironments in part by the “education” of immune cells attracted via chemotactic axes such as that of CCR5-CCL5. Further, CCR5 upregulation by cancer cells, coupled with its association with pro-tumorigenic features such as drug resistance and metastasis, has suggested CCR5 as a therapeutic target. However, with several conformational “pools” being reported, phenotypic investigations must be capable of unveiling conformational heterogeneity. Addressing this challenge, we performed super-resolution structured illumination microscopy (SIM) and single molecule partially TIRF-coupled HILO (PaTCH) microscopy of CCR5 in fixed cells. SIM data revealed a non-random spatial distribution of CCR5 assemblies, while Intensity-tracking of CCR5 assemblies from PaTCH images indicated dimeric sub-units independent of CCL5 perturbation. These biophysical methods can provide important insights into the structure and function of onco-immunogenic receptors and many other biomolecules. SIM and PaTCH used for precise bioimaging and single-molecule tracking of CCR5 Super-resolved SIM shows non-random CCR5 clustering Single-molecule PaTCH reveals molecular stoichiometry of CCR5 assemblies Immunology; Biophysics; Cancer
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影响因子：
5.8
作者：
Lenn T;Leake MC
通讯作者：
Leake MC
筏结构域的动态重新分布是细胞趋化过程中信号传导的组织平台。
影响因子：
7.8
作者：
Gómez-Moutón, C;Lacalle, RA;Mira, E;Jiménez-Baranda, S;Barber, DF;Carrera, AC;Martínez, C;Mañes, S
通讯作者：
Mañes, S
影响因子：
6.7
作者：
Colin, Philippe;Zhou, Zhicheng;Lagane, Bernard
通讯作者：
Lagane, Bernard
影响因子：
11.2
作者：
Chang, Li-Yuan;Lin, Yung-Chang;Lin, Chun-Yen
通讯作者：
Lin, Chun-Yen
DOI：
10.1016/j.ymeth.2020.06.007
发表时间：
2021-09
期刊：
Methods (San Diego, Calif.)
影响因子：
--
作者：
Dresser L;Hunter P;Yendybayeva F;Hargreaves AL;Howard JAL;Evans GJO;Leake MC;Quinn SD
通讯作者：
Quinn SD