West J W, Patton D E, Scheuer T, Wang Y, Goldin A L, Catterall W A
Department of Pharmacology, University of Washington, Seattle 98195.
Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10910-4. doi: 10.1073/pnas.89.22.10910.
The inward Na+ current underlying the action potential in nerve is terminated by inactivation. The preceding report shows that deletions within the intracellular linker between domains III and IV remove inactivation, but mutation of conserved basic and paired acidic amino acids has little effect. Here we show that substitution of glutamine for three clustered hydrophobic amino acids, Ile-1488, Phe-1489, and Met-1490, completely removes fast inactivation. Substitution of Met-1490 alone slows inactivation significantly, substitution of Ile-1488 alone both slows inactivation and makes it incomplete, and substitution of Phe-1489 alone removes inactivation nearly completely. These results demonstrate an essential role of Phe-1489 in Na(+)-channel inactivation. It is proposed that the hydrophobic cluster of Ile-1488, Phe-1489, and Met-1490 serves as a hydrophobic latch that stabilizes the inactivated state in a hinged-lid mechanism of Na(+)-channel inactivation.
神经动作电位所依赖的内向Na⁺电流通过失活而终止。之前的报告显示,结构域III和IV之间的细胞内连接区的缺失会消除失活,但保守的碱性和成对酸性氨基酸的突变影响很小。在此我们表明,用谷氨酰胺取代三个成簇的疏水氨基酸Ile-1488、Phe-1489和Met-1490,会完全消除快速失活。单独取代Met-1490会显著减慢失活,单独取代Ile-1488会使失活减慢且不完全,而单独取代Phe-1489几乎能完全消除失活。这些结果证明了Phe-1489在Na⁺通道失活中起关键作用。有人提出,Ile-1488、Phe-1489和Met-1490的疏水簇作为一个疏水锁扣,以Na⁺通道失活的铰链盖机制稳定失活状态。