Abstract
The sodium (Na + )-calcium (Ca 2+ ) exchanger 1 (NCX1) is an antiporter membrane protein encoded by the SLC8A1 gene. In the heart, it maintains cytosolic Ca 2+ homeostasis, serving as the primary mechanism for Ca 2+ extrusion during relaxation. Dysregulation of NCX1 is observed in end-stage human heart failure. In this study we used affinity purification coupled with mass spectrometry in rat left ventricle lysates to identify novel NCX1 interacting proteins in the heart. Two screens were conducted using: 1) anti-NCX1 against endogenous NCX1 and 2) anti-His with His-TF-NCX1 cyt recombinant protein as bait. The respective methods identified 112 and 350 protein partners, of which several were known NCX1 partners from the literature and 29 occurred in both screens. Selected protein partners were validated for binding to NCX1 expressed in HEK293 cells. A cardiac NCX1 protein-protein interaction map was constructed. The map was highly connected, containing distinct clusters of proteins with different biological functions, where cell communication and signal transduction formed the largest clusters. The NCX1 interactome was also significantly enriched with proteins/genes involved in cardiovascular disease. Exploring the molecular mechanisms of these protein partners in future studies may aid in elucidation of NCX1 regulation and facilitate selective therapeutic targeting of NCX1.
Published Version
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call