Direct detection of SERCA calcium transport and small-molecule inhibition in giant unilamellar vesicles.

ABSTRACT: We have developed a charge-mediated fusion method to reconstitute the sarco/endoplasmic reticulum Ca²⁺-ATPase (SERCA) in giant unilamellar vesicles (GUV). Intracellular Ca²⁺ transport by SERCA controls key processes in human cells such as proliferation, signaling, and contraction. Small-molecule effectors of SERCA are urgently needed as therapeutics for Ca²⁺ dysregulation in human diseases including cancer, diabetes, and heart failure. Here we report the development of a method for efficiently reconstituting SERCA in GUV, and we describe a streamlined protocol based on optimized parameters (e.g., lipid components, SERCA preparation, and activity assay requirements). ATP-dependent Ca²⁺ transport by SERCA in single GUV was detected directly using c

SUBMITTER: Bian T 

PROVIDER: S-EPMC5123963 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature