Redistribution of SERCA calcium pump conformers during intracellular calcium signaling

The Journal of Biological Chemistry
Olga N RaguimovaSeth L Robia

Abstract

The conformational changes of a calcium transport ATPase were investigated with molecular dynamics (MD) simulations as well as fluorescence resonance energy transfer (FRET) measurements to determine the significance of a discrete structural element for regulation of the conformational dynamics of the transport cycle. Previous MD simulations indicated that a loop in the cytosolic domain of the SERCA calcium transporter facilitates an open-to-closed structural transition. To investigate the significance of this structural element, we performed additional MD simulations and new biophysical measurements of SERCA structure and function. Rationally designed in silico mutations of three acidic residues of the loop decreased SERCA domain-domain contacts and increased domain-domain separation distances. Principal component analysis of MD simulations suggested decreased sampling of compact conformations upon N-loop mutagenesis. Deficits in headpiece structural dynamics were also detected by measuring intramolecular FRET of a Cer-YFP-SERCA construct (2-color SERCA). Compared with WT, the mutated 2-color SERCA shows a partial FRET response to calcium, whereas retaining full responsiveness to the inhibitor thapsigargin. Functional measureme...Continue Reading

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Citations

Sep 11, 2019·Cold Spring Harbor Perspectives in Biology·Jialin ChenPeter Vangheluwe
Nov 2, 2019·Biochemical Society Transactions·Mateusz DylaMagnus Kjaergaard
Dec 25, 2019·Annual Review of Biochemistry·Mateusz DylaPoul Nissen
May 14, 2020·International Journal of Molecular Sciences·Bela PappHoma Adle-Biassette
Mar 23, 2019·American Journal of Physiology. Heart and Circulatory Physiology·Elisa BovoAleksey V Zima
Jun 14, 2020·International Journal of Molecular Sciences·Rodrigo Aguayo-Ortiz, L Michel Espinoza-Fonseca
Oct 7, 2020·International Journal of Molecular Sciences·Rodrigo Aguayo-Ortiz, L Michel Espinoza-Fonseca
Apr 8, 2020·Computational and Structural Biotechnology Journal·Eli Fernández-de GortariL Michel Espinoza-Fonseca
Oct 19, 2020·Biophysical Journal·Olga N RaguimovaL Michel Espinoza-Fonseca

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