Jennie Sjöhamn1, Kristina Hedfalk2. 1. Department of Chemistry and Molecular Biology, University of Gothenburg, P.O. Box 462, SE-405 30 Göteborg, Sweden. 2. Department of Chemistry and Molecular Biology, University of Gothenburg, P.O. Box 462, SE-405 30 Göteborg, Sweden. Electronic address: kristina.hedfalk@chem.gu.se.
Abstract
BACKGROUND: Insight into protein-protein interactions (PPIs) is highly desirable in order to understand the physiology of cellular events. This understanding is one of the challenges in biochemistry and molecular biology today, especially for eukaryotic membrane proteins where hurdles of production, purification and structural determination must be passed. SCOPE OF REVIEW: We have explored the common strategies used to find medically relevant interaction partners of aquaporins (AQPs). The most frequently used methods to detect direct contact, yeast two-hybrid interaction assay and co-precipitation, are described together with interactions specifically found for the selected targets AQP0, AQP2, AQP4 and AQP5. MAJOR CONCLUSIONS: The vast majority of interactions involve the aquaporin C-terminus and the characteristics of the interaction partners are strikingly diverse. While the well-established methods for PPIs are robust, a novel approach like bimolecular fluorescence complementation (BiFC) is attractive for screening many conditions as well as transient interactions. The ultimate goal is structural evaluation of protein complexes in order to get mechanistic insight into how proteins communicate at a molecular level. GENERAL SIGNIFICANCE: What we learn from the human aquaporin field in terms of method development and communication between proteins can be of major use for any integral membrane protein of eukaryotic origin. This article is part of a Special Issue entitled Aquaporins.
BACKGROUND: Insight into protein-protein interactions (PPIs) is highly desirable in order to understand the physiology of cellular events. This understanding is one of the challenges in biochemistry and molecular biology today, especially for eukaryotic membrane proteins where hurdles of production, purification and structural determination must be passed. SCOPE OF REVIEW: We have explored the common strategies used to find medically relevant interaction partners of aquaporins (AQPs). The most frequently used methods to detect direct contact, yeast two-hybrid interaction assay and co-precipitation, are described together with interactions specifically found for the selected targets AQP0, AQP2, AQP4 and AQP5. MAJOR CONCLUSIONS: The vast majority of interactions involve the aquaporin C-terminus and the characteristics of the interaction partners are strikingly diverse. While the well-established methods for PPIs are robust, a novel approach like bimolecular fluorescence complementation (BiFC) is attractive for screening many conditions as well as transient interactions. The ultimate goal is structural evaluation of protein complexes in order to get mechanistic insight into how proteins communicate at a molecular level. GENERAL SIGNIFICANCE: What we learn from the human aquaporin field in terms of method development and communication between proteins can be of major use for any integral membrane protein of eukaryotic origin. This article is part of a Special Issue entitled Aquaporins.
Authors: Susanna Törnroth-Horsefield; Clara Chivasso; Helin Strandberg; Claudia D'Agostino; Carla V T O'Neale; Kevin L Schey; Christine Delporte Journal: Int J Mol Sci Date: 2022-08-25 Impact factor: 6.208