Transmembrane interactions in the activation of the Neu receptor tyrosine kinase.
Transmembrane interactions in the activation of the Neu receptor tyrosine kinase.
复制标题
Neu 受体酪氨酸激酶激活中的跨膜相互作用。
DOI：
10.1021/bi012117l
复制
发表时间：
2002
期刊：
影响因子：
2.9
通讯作者：
Aimoto,Saburo
中科院分区：
文献类型：
--
作者：
Smith,StevenO;Smith,Charles;Shekar,Srinivasan;Peersen,Olve;Ziliox,Martine;Aimoto,Saburo
Neu受体酪氨酸激酶通过受体跨膜结构域中的单个氨基酸变化而组成型激活。Val 664突变为谷氨酸或谷氨酰胺诱导受体二聚化和受体胞内激酶结构域的自磷酸化。这种单一突变激活受体的能力是序列依赖性的，表明特定的螺旋-螺旋相互作用稳定了跨膜二聚体。我们已经确定了本地的二级结构和螺旋间的接触在该区域的位置664肽模型的激活受体使用固态旋转共振和旋转回波双共振（REDOR）NMR方法。在对应于野生型Neu和激活的Neu* 跨膜序列的肽上的1- 13 C-Thr 662和2- 13 C-Gly 665之间进行螺旋内13 C旋转共振距离测量，所述激活的Neu* 跨膜序列分别在位置664处含有缬氨酸和谷氨酸。我们在Neu和Neu* 中观察到相似的核间距（4.5 ± 0.2 μ m），表明残基664附近的区域是螺旋形的，不受突变的影响。相对螺旋上Gln 664侧链之间的螺旋间15 N···13 C REDOR测量与侧链官能团之间的氢键不一致。然而，1- 13 C-Glu 664和2- 13 C-Gly 665之间以及1- 13 C-Gly 665和2- 13 C-Gly 665之间的螺旋间旋转共振测量表明紧密接触（4.3 - 4.5 Ω），与Gly 665在Neu* 二聚体界面中的堆积一致。这些测量为跨膜二聚体建模提供了结构约束，并定义了活化受体中跨膜螺旋的旋转方向。
The Neu receptor tyrosine kinase is constitutively activated by a single amino acid change in the transmembrane domain of the receptor. The mutation of Val664 to glutamate or glutamine induces receptor dimerization and autophosphorylation of the receptor's intracellular kinase domain. The ability of this single mutation to activate the receptor is sequence-dependent, suggesting that specific helix−helix interactions stabilize the transmembrane dimer. We have determined the local secondary structure and interhelical contacts in the region of position 664 in peptide models of the activated receptor using solid-state rotational resonance and rotational echo double-resonance (REDOR) NMR methods. Intrahelical13C rotational resonance distance measurements were made between 1-13C-Thr662 and 2-13C-Gly665 on peptides corresponding to the wild-type Neu and activated Neu* transmembrane sequences containing valine and glutamate at position 664, respectively. We observed similar internuclear distances (4.5 ± 0.2 Å) in both Neu and Neu*, indicating that the region near residue 664 is helical and is not influenced by mutation. Interhelical15N···13C REDOR measurements between Gln664 side chains on opposing helices were not consistent with hydrogen bonding between the side chain functional groups. However, interhelical rotational resonance measurements between 1-13C-Glu664 and 2-13C-Gly665 and between 1-13C-Gly665 and 2-13C-Gly665 demonstrated close contacts (4.3−4.5 Å) consistent with the packing of Gly665 in the Neu* dimer interface. These measurements provide structural constraints for modeling the transmembrane dimer and define the rotational orientation of the transmembrane helices in the activated receptor.