Biochemical and NMR studies reveal specific interaction between STIMATE C-tail and PI(4,5)P2 or PI(3,4,5)P3-containing membrane

Chongxu Liu; Youjia Zhang; Liang Ge; Ling Li; Bo Wu; Junfeng Wang

doi:10.1016/j.bbrc.2022.01.100

Biochemical and NMR studies reveal specific interaction between STIMATE C-tail and PI(4,5)P₂ or PI(3,4,5)P₃-containing membrane

Biochem Biophys Res Commun. 2022 Mar 15:597:16-22. doi: 10.1016/j.bbrc.2022.01.100. Epub 2022 Jan 29.

Authors

Chongxu Liu¹, Youjia Zhang¹, Liang Ge², Ling Li¹, Bo Wu³, Junfeng Wang⁴

Affiliations

¹ High Magnetic Field Laboratory, Key Laboratory of High Magnetic Field and Ion Beam Physical Biology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei, Anhui, 230031, PR China; University of Science and Technology of China, Hefei, Anhui, 230036, PR China.
² High Magnetic Field Laboratory, Key Laboratory of High Magnetic Field and Ion Beam Physical Biology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei, Anhui, 230031, PR China.
³ High Magnetic Field Laboratory, Key Laboratory of High Magnetic Field and Ion Beam Physical Biology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei, Anhui, 230031, PR China. Electronic address: wubo@hmfl.ac.cn.
⁴ High Magnetic Field Laboratory, Key Laboratory of High Magnetic Field and Ion Beam Physical Biology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei, Anhui, 230031, PR China; Institutes of Physical Science and Information Technology, Anhui University, Hefei, Anhui, 230601, PR China. Electronic address: junfeng@hmfl.ac.cn.

PMID: 35121178
DOI: 10.1016/j.bbrc.2022.01.100

Abstract

STIMATE is an endoplasmic reticulum (ER) resident membrane protein that plays key roles in regulating calcium signaling occurring at ER-plasma membrane (PM) junctions. It is also involved in the regulation of ER-PM junction maintenance. STIMATE contains multiple putative transmembrane domains with a polybasic C tail (STIMATE-CT) that directly interacts with stromal interaction molecule 1 (STIM1) to promote STIM1 conformational switch. Here using liposome pulldown assay, we show that STIMATE-CT can specifically interact with PI(4,5)P₂ or PI(3,4,5)P₃-containing membrane. NMR analysis indicates that STIMATE-CT is intrinsically disordered. Furthermore, NMR titration with bicelles and mutation analysis reveal that the regions of ²⁴²VRYR²⁴⁵ and ²⁸⁴KKKK²⁸⁷ in STIMATE-CT are both essential for its membrane binding.

Keywords: Intrinsically disordered; Membrane; NMR; PI(3,4,5)P(3); PI(4,5)P(2); STIMATE C-tail.