Comparison of nerve growth factor receptor binding models using heterodimeric muteins.
Comparison of nerve growth factor receptor binding models using heterodimeric muteins.
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使用异二聚体静脉素对神经生长因子受体结合模型的比较。
DOI：
10.1002/jnr.23116
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发表时间：
2012-12
影响因子：
4.2
通讯作者：
Neet, Kenneth E.
中科院分区：
文献类型：
--
作者：
Mehta, Hrishikesh M.;Woo, Sang B.;Neet, Kenneth E.
神经生长因子(NGF)是一种同源二聚体，可与两种不同类型的受体TrkA和p75结合，以支持神经元的生存和分化。细胞表面的高亲和力结合被认为涉及一个异型受体复合体，但其确切性质尚不清楚。我们开发了一种由两个NGF突变体组成的异源二聚体(Heteromutein)，它可以与异源二聚体两侧的p75和TrkA结合，但不能结合两个TrkA受体。以前描述的突变体是TrkA阴性的Δ9/13和通过TrkA信号选择性的7-84-103。在假定没有NGF诱导TrkA二聚的情况下，异型肌动蛋白(Htm1)用于研究异型受体复合体的形成和功能。细胞结合分析表明，Htm1与TrkA的结合不如野生型(Wt)NGF有效，但亲和力强于任何一种同源二聚体突变体。Htm1、7-84-103和Δ9/13都能够竞争与PC12细胞的低温、冷追稳定结合，这表明这种高亲和力结合需要一部分与p75结合。与wtNGF相比，htm1在PC12细胞中的存活、突起生长和MAPK信号转导也表现出降低的反应，但在wtNGF>；htm1>；7-84-103>；Δ9/13的顺序上好于亲本突变体。在关于NGF受体结合机制的长期争论中，我们的数据支持NGF的配体从p75传递到TrkA，涉及p75-NGF-TrkA的瞬时异位受体复合体。
Nerve growth factor (NGF) is a homodimer that binds to two distinct receptor types, TrkA and p75, to support survival and differentiation of neurons. The high-affinity binding on the cell surface is believed to involve a heteroreceptor complex, but its exact nature is unclear. We developed a heterodimer (heteromutein) of two NGF muteins that can bind p75 and TrkA on opposite sides of the heterodimer, but not two TrkA receptors. Previously described muteins are Δ9/13 that is TrkA negative and 7-84-103 that is signal selective through TrkA. The heteromutein (Htm1) was used to study the heteroreceptor complex formation and function, in the putative absence of NGF-induced TrkA dimerization. Cellular binding assays indicated that Htm1 does not bind TrkA as efficiently as wild-type (wt) NGF but has better affinity than either homodimeric mutein. Htm1, 7-84-103, and Δ9/13 were each able to compete for cold-temperature, cold-chase stable binding on PC12 cells, indicating that binding to p75 was required for a portion of this high-affinity binding. Survival, neurite outgrowth, and MAPK signaling in PC12 cells also showed a reduced response for Htm1, compared with wtNGF, but was better than the parent muteins in the order wtNGF > Htm1 > 7-84-103 >> Δ9/13. Htm1 and 7-84-103 demonstrated similar levels of survival on cells expressing only TrkA. In the longstanding debate on the NGF receptor binding mechanism, our data support the ligand passing of NGF from p75 to TrkA involving a transient heteroreceptor complex of p75-NGF-TrkA.
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DOI：
10.1016/j.bbagen.2010.06.007
发表时间：
2010-09-01
影响因子：
3
作者：
Brahimi, Fouad;Liu, Jing;Saragovi, H. Uri
通讯作者：
Saragovi, H. Uri
影响因子：
4.7
作者：
BURTON, LE;SCHMELZER, CH;GORRELL, A
通讯作者：
GORRELL, A
影响因子：
4.8
作者：
Lad, SP;Peterson, DA;Neet, KE
通讯作者：
Neet, KE
影响因子：
64.8
作者：
Gong, Yong;Cao, Peng;Jiang, Tao
通讯作者：
Jiang, Tao
DOI：
10.1124/jpet.104.066225
发表时间：
2004-08-01
影响因子：
3.5
作者：
Colquhoun, A;Lawrance, GM;Ross, GM
通讯作者：
Ross, GM