FERM domain of moesin desorbs the basic-rich cytoplasmic domain of l-selectin from the anionic membrane surface.

FERM domain of moesin desorbs the basic-rich cytoplasmic domain of l-selectin from the anionic membrane surface.
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DOI:
10.1016/j.jmb.2013.06.008
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发表时间:
2013-09-23
影响因子:
5.6
通讯作者:
Li, Renhao
Li, Renhao
中科院分区:
生物学2区
文献类型:
--
作者:
Deng, Wei;Cho, Sungyun;Li, Renhao

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Moesin 和钙调蛋白 (CaM) 共同与细胞中 L-选择素的胞质结构域结合,调节 L-选择素的功能和胞外域脱落。使用荧光光谱,我们检查了 moesin FERM 结构域与在模型磷脂脂质体中重建的 L-选择素 (CLS) 的重组跨膜和细胞质结构域的关联。磷脂酰胆碱脂质体中moesin FERM结构域与CLS的解离常数约为300 nM。与破坏 CaM 与 CLS 的关联相反,磷脂酰胆碱脂质体中包含阴离子磷脂酰丝氨酸脂质增加了 moesin FERM 结构域对 CLS 的表观结合亲和力。利用附着在 CLS 细胞质结构域上的环境敏感荧光探针和附着在脂双层上的硝基氧猝灭剂,我们发现 moesin FERM 结构域的结合诱导了 CLS 富含碱性的细胞质结构域从阴离子膜表面的解吸,从而使 CaM 随后与 CLS 的细胞质结构域结合。这些结果阐明了 moesin/l-选择素/CaM 三元复合物的分子基础,并表明磷脂在调节 l-选择素功能和脱落中的重要作用。
Moesin and calmodulin (CaM) jointly associate with the cytoplasmic domain of l-selectin in the cell to modulate the function and ectodomain shedding of l-selectin. Using fluorescence spectroscopy, we have examined the association of moesin FERM domain with the recombinant transmembrane and cytoplasmic domains of l-selectin (CLS) reconstituted in model phospholipid liposomes. The dissociation constant of moesin FERM domain to CLS in the phosphatidylcholine liposome is about 300 nM. In contrast to disrupting the CaM association with CLS, inclusion of anionic phosphatidylserine lipids in the phosphatidylcholine liposome increased the apparent binding affinity of moesin FERM domain for CLS. Using the environmentally sensitive fluorescent probe attached to the cytoplasmic domain of CLS and the nitroxide quencher attached to the lipid bilayer, we showed that the association of moesin FERM domain induced the desorption of the basic-rich cytoplasmic domain of CLS from the anionic membrane surface, which enabled subsequent association of CaM to the cytoplasmic domain of CLS. These results have elucidated the molecular basis for the moesin/l-selectin/CaM ternary complex and suggested an important role of phospholipids in modulating l-selectin function and shedding.
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