Primitive Phospholamban- and Sarcolipin-like Peptides Inhibit the Sarcoplasmic Reticulum Calcium Pump SERCA

• Published in: Biochemistry (Easton) Vol. 61; no. 14; pp. 1419 - 1430
• Main Authors: Bak, Jessi J., Aguayo-Ortiz, Rodrigo, Rathod, Nishadh, Primeau, Joseph O., Khan, Muhammad Bashir, Robia, Seth L., Lemieux, M. Joanne, Espinoza-Fonseca, L. Michel, Young, Howard S.
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 19.07.2022
• Subjects: Animals; Calcium - metabolism; Calcium Signaling; Calcium-Binding Proteins - chemistry; Humans; Mammals - metabolism; Molecular Dynamics Simulation; Muscle Proteins; Peptides - metabolism; Peptides - pharmacology; Proteolipids - chemistry; Sarcoplasmic Reticulum - metabolism; Sarcoplasmic Reticulum Calcium-Transporting ATPases - chemistry
• ISSN: 0006-2960; 1520-4995; 1520-4995
• DOI: 10.1021/acs.biochem.2c00246

Intracellular calcium signaling is essential for all kingdoms of life. An important part of this process is the sarco-endoplasmic reticulum Ca2+-ATPase (SERCA), which maintains the low cytosolic calcium levels required for intracellular calcium homeostasis. In higher organisms, SERCA is regulated by a series of tissue-specific transmembrane subunits such as phospholamban in cardiac muscles and sarcolipin in skeletal muscles. These regulatory axes are so important for muscle contractility that SERCA, phospholamban, and sarcolipin are practically invariant across mammalian species. With the recent discovery of the arthropod sarcolambans, the family of calcium pump regulatory subunits appears to span more than 550 million years of evolutionary divergence from arthropods to humans. This evolutionary divergence is reflected in the peptide sequences, which vary enormously from one another and only vaguely resemble phospholamban and sarcolipin. The discovery of the sarcolambans allowed us to address two questions. How much sequence variation is tolerated in the regulation of mammalian SERCA activity by the transmembrane peptides? Do divergent peptide sequences mimic phospholamban or sarcolipin in their regulatory activities despite limited sequence similarity? We expressed and purified recombinant sarcolamban peptides from three different arthropods. The peptides were coreconstituted into proteoliposomes with mammalian SERCA1a and the effect of each peptide on the apparent calcium affinity and maximal activity of SERCA was measured. All three peptides were superinhibitors of SERCA, exhibiting either phospholamban-like or sarcolipin-like characteristics. Molecular modeling, protein–protein docking, and molecular dynamics simulations revealed novel features of the divergent peptides and their SERCA regulatory properties.