ACTA, a fluorescent analogue of thapsigargin, is a potent inhibitor and a conformational probe of skeletal muscle Ca2+‐ATPase

FEBS Letters

Brøgger Christensen, Søren¹  Procida, Kristina²  Kromann, Hasse¹  Treiman, Marek²  Caspersen, Casper² 

[1] Institute of Medicinal Chemistry, The Royal Danish School of Pharmacy, Universitetsparken 2, DK-2100 Copenhagen Ø, Denmark;Department of Medical Physiology–Center for Cellular Communication, University of Copenhagen, The Panum Institute, 12.2, Blegdamsvej 3C, DK-2200 Copenhagen N, Denmark

关键词: Ca2+ -ATPase;    Sarco-endoplasmic reticulum Ca2+-ATPase pump;    Calcium;    Fluorescence;    Thapsigargin;    DMSO;    dimethylsulfoxide;    ER;    endoplasmic reticulum;    SDS;    sodium dodecyl sulfate;    SERCA;    sarco-endoplasmic reticulum Ca2+-ATPase;    SR;    sarcoplasmic reticulum;    TES;    N-tris(hydroxymethyl)-methyl-2-aminomethane sulfonic acid;   

DOI  :  10.1016/S0014-5793(98)01352-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Thapsigargin is a highly potent and selective inhibitor of sarco-endoplasmic reticulum (SERCA) family of Ca²⁺-ATPases and a useful tool in research concerning the function of intracellular Ca²⁺ stores. We describe here a novel fluorescent derivative (8-O-(4-aminocinnamoyl)-8-O-debutanoylthapsigargin, termed ACTA) of this compound, acting as a Ca²⁺-ATPase inhibitor with a potency approaching that of thapsigargin. Binding of ACTA to the skeletal muscle sarcoplasmic reticulum vesicles results in a strong fluorescence enhancement, approximately 66% of which depends on ACTA association with Ca²⁺-ATPase. This specific component of ACTA fluorescence is sensitive to the E₁-E₂ conformational equilibrium of the pump. The combined properties of high potency and binding-dependent fluorescence suggest ACTA to be a useful probe for a range of studies involving the SERCA class of ATPases.

【 授权许可】

Unknown