TRPV4 expresses in bone cell lineages and TRPV4-R616Q mutant causing Brachyolmia in human reveals “loss-of-interaction” with cholesterol

• Published in: Biochemical and biophysical research communications 2019-10, Vol.517 (4), p.566-574
• Main Authors: Das, Rashmita, Goswami, Chandan
• Format: Article
• Language: English
• Subjects: Channelopathy; Cholesterol; Lipid-protein interaction; Mesenchymal stem cells; Osteogenesis; TRP channels
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2019.07.042

Transient receptor potential Vanilloid ion channel sub type 4 (TRPV4) is involved in complex Ca2+-signaling. At least one copy of TRPV4 is present in all vertebrates and is involved in several physiological processes including sensory process and point mutations in TRPV4 leads to development of different pathophysiological disorders in human. R616Q mutant of TRPV4 has been referred as “gain-of-function” mutant causing abnormality in bone cells and develop pathophysiological condition known as “Brachyolmia”. In this work, we demonstrated that R616Q mutation is located in a very critical position of TRPV4 containing a cholesterol-binding motif sequence which is highly conserved in all vertebrates. Accordingly, TRPV4-Wt but not the TRPV4-R616Q localizes preferably in cholesterol-enriched lipid rafts in osteogenic cell line Saos2 and in DRG-neuron derived F11 cell line. Further, FRAP experiment suggest TRPV4-Wt but not the TRPV4-R616Q mutant is more mobile especially in cholesterol-reduced lipid membrane. GST-tagged TM4-Loop-TM5 fragment containing TRPV4-Wt but not R616Q sequence interacts with cholesterol, forms high-molecular weight complex and also show band shift in SDS-PAGE. TRPV4 is expressed in Mesenchymal stem cells and the localization of TRPV4 in lipid raft is dependent on temperature and cholesterol. Our data suggests that TRPV4-R616Q mutant behaves as a “loss-of-interaction” with cholesterol. •TRPV4 possesses several cholesterol binding motifs which are conserved throughout vertebrate evolution.•TRPV4-Wt but R616Q mutant binds to cholesterol and forms higher molecular complex.•TRPV4-Wt but not R616Q mutant localizes in the lipid raft.•The “loss-of-interaction” of cholesterol in case of R-616Q mutant is suggestive for the regulatory role of cholesterol in case of TRPV4 function.•The “loss-of-interaction” with cholesterol correlates with the “gain-of-function” for TRPV4.