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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).
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http://dx.doi.org/10.1016/j.xpro.2022.101316 | DOI Listing |
J Biol Chem
September 2025
Department of Biochemistry, Stanford University School of Medicine, Stanford, CA, United States; Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, San Francisco, CA, United States. Electronic address:
PPM1H phosphatase reverses Parkinson's disease-associated, Leucine Rich Repeat Kinase 2-mediated, Rab GTPase phosphorylation. We showed previously that PPM1H relies on an N-terminal amphipathic helix for Golgi membrane localization and this helix enables PPM1H to associate with liposomes in vitro; binding to highly curved liposomes activates PPM1H's phosphatase activity. We show here that PPM1H also contains an allosteric binding site for its non-phosphorylated reaction products, Rab8A and Rab10.
View Article and Find Full Text PDFJ Biol Chem
September 2025
Department of Biology, University of Fribourg, Chemin du Musée 10, 1700 Fribourg, Switzerland. Electronic address:
Cellular membranes maintain distinct lipid compositions, with sterols enriched in the plasma membrane despite their synthesis in the endoplasmic reticulum (ER). This distribution relies on vesicular and non-vesicular transport, the latter facilitated by lipid transfer proteins (LTPs) at membrane contact sites. In yeast, the Lam/Ltc family of LTPs is critical for sterol transport.
View Article and Find Full Text PDFFront Immunol
September 2025
College of Pharmacy, Liaoning University of Traditional Chinese Medicine, Dalian, China.
Background: Colorectal cancer (CRC) remains a leading cause of cancer-related mortality worldwide, with pro-tumor inflammation playing a critical role in its initiation and progression. Chronic inflammation acts as a major driving force and a distinct mechanism underlying tumorigenesis. Although previous studies have demonstrated the importance of the VEGF/p38MAPK and p38MAPK/HSP27 signaling pathways in CRC-associated inflammation, a comprehensive understanding of the entire pro-tumor inflammatory mechanism remains incomplete.
View Article and Find Full Text PDFTrends Plant Sci
August 2025
School of Life Science, Yangtze University, Jingzhou, China. Electronic address:
Protein Sci
September 2025
School of Pharmaceutical Sciences, Faculty of Medicine, University of Ottawa, Ottawa, Ontario, Canada.
The nucleus, as the control center of the eukaryotic cell, is a prime target for therapeutic interventions due to its role in regulating genetic material. Importin-α is critical for successful nuclear import as it recognizes and binds to cargo proteins bearing a classical nuclear localization signal (NLS), which facilitates their transport from the cytoplasm into the nucleus. NLS tagging to 'actively' import therapeutics provides the most effective means of maximizing nuclear localization and therapeutic efficacy.
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