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 Agric Food Chem
March 2025
Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu 96822, Hawaii United States.
Autophagy-associated protein 8 (ATG8) is essential for autophagy and organismal growth and development. In this study, we successfully resolved the crystal structure of () ATG8a (ATG8a) at 1.36 Å resolution.
View Article and Find Full Text PDFHortic Res
March 2025
State Key Laboratory of Tree Genetics and Breeding, Research Institute of Tropical Forestry, Chinese Academy of Forestry, 682 Guangshan Road, Tianhe District, Guangzhou 510520, Guangdong, China.
Sandalwood (), a culturally significant and economically valuable horticultural species, is renowned for its heartwood and essential oils enriched with sesquiterpene compounds such as santalol. Despite progress in elucidating the biosynthetic pathway of these valuable metabolites, the transcriptional regulation of this process, particularly under abiotic stress conditions, remains largely unexplored. Under drought conditions, we observed a marked increase in expression, paralleled by elevated levels of santalols.
View Article and Find Full Text PDFBiophys Rev
February 2025
Institute of Biochemistry and Biophysics PAS, Pawinskiego 5a, 02-106 Warsaw, Poland.
Various biophysical and biochemical techniques have been developed to measure the affinity of interacting molecules. This review analyzes the combination of three methods: differential scanning fluorimetry as the initial high-throughput screening technique and microscale thermophoresis and isothermal titration calorimetry as complementary methods to quantify binding affinity. The presented work is the first to detailed compare the strengths and flaws of these three specific methods, as well as their application possibilities and complementarity.
View Article and Find Full Text PDFMol Immunol
March 2025
Department of Experimental Medical Science, Lund University, BMC C13, Lund 22184, Sweden; LINXS - Institute of Advanced Neutron and X-ray Science, Scheelevägen 19, Lund 223 70, Sweden. Electronic address:
Bacterial toxins, called superantigens, are produced by Staphylococcus aureus and are known to activate γδ T cells. γδ T cells contribute to long-lasting immunity against bacterial skin infections caused by S. aureus.
View Article and Find Full Text PDFbioRxiv
February 2025
Department of Biological Chemistry, University of California, Los Angeles, 615 Charles E. Young Drive South, Los Angeles, California 90095, United States.
Cocktail soaking using single-crystal X-ray diffraction (SC-XRD) has previously allowed high-throughput crystallographic screening of ligands against protein targets. However, protein microcrystals are not amenable to this approach if they are too small to yield strong diffraction patterns. In this study, we developed a workflow integrating cocktail soaking with automated microcrystal electron diffraction (MicroED) to allow rapid ligand screening, structure determination, and binding analysis directly from microcrystals.
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