Characterization of human aquaporin protein-protein interactions using microscale thermophoresis (MST)

STAR Protoc. 2022 Apr 15;3(2):101316. doi: 10.1016/j.xpro.2022.101316. eCollection 2022 Jun 17.

Abstract

Aquaporin water channels (AQPs) are membrane proteins that maintain cellular water homeostasis. The interactions between human AQPs and other proteins play crucial roles in AQP regulation by both gating and trafficking. Here, we describe a protocol for characterizing the interaction between a human AQP and a soluble interaction partner using microscale thermophoresis (MST). MST has the advantage of low sample consumption and high detergent compatibility enabling AQP protein-protein interaction investigation with a high level of control of components and environment. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020) and Roche et al. (2017).

Keywords: Cell Membrane; Protein Biochemistry; Single-molecule Assays.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aquaporins* / metabolism
  • Homeostasis
  • Humans
  • Proteins / metabolism

Substances

  • Aquaporins
  • Proteins