A kink in DWORF helical structure controls the activation of the sarcoplasmic reticulum Ca2+-ATPase

U. Venkateswara Reddy; Daniel K Weber; Songlin Wang; Erik Larsen; Alfonso De Simone; Gopinath Tata; Seth Robia; Gianluigi Veglia

doi:10.1016/j.str.2021.11.003

A kink in DWORF helical structure controls the activation of the sarcoplasmic reticulum Ca²⁺-ATPase

U. Venkateswara Reddy, Daniel K Weber, Songlin Wang, Erik Larsen, Alfonso De Simone, Gopinath Tata, Seth Robia, Gianluigi Veglia

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

SERCA is a P-type ATPase embedded in the sarcoplasmic reticulum and plays a central role in muscle relaxation. SERCA's function is regulated by single-pass membrane proteins called regulins. Unlike other regulins, dwarf open reading frame (DWORF) expressed in cardiac muscle has a unique activating effect. Here, we determine the structure and topology of DWORF in lipid bilayers using a combination of oriented sample solid-state NMR spectroscopy and replica-averaged orientationally restrained molecular dynamics. We found that DWORF's structural topology consists of a dynamic N-terminal domain, an amphipathic juxtamembrane helix that crosses the lipid groups at an angle of 64°, and a transmembrane C-terminal helix with an angle of 32°. A kink induced by Pro15, unique to DWORF, separates the two helical domains. A single Pro15Ala mutant significantly decreases the kink and eliminates DWORF's activating effect on SERCA. Overall, our findings directly link DWORF's structural topology to its activating effect on SERCA.

Original language	English (US)
Pages (from-to)	360-370.e6
Journal	Structure
Volume	30
Issue number	3
DOIs	https://doi.org/10.1016/j.str.2021.11.003
State	Published - Mar 3 2022

Bibliographical note

Funding Information:
D.K.W. was supported by an American Heart Association postdoctoral fellowship (19POST34420009). This work was supported by National Heart, Lung, and Blood Institute grants HL143816 to S.R. and G.V. and HL092321 to S.R. We thank Caitlin Walker for her assistance with cloning the DWORF P15A expression construct and Dr. Maximo Sanz-Hernández for his contributions to the modeling work.

Publisher Copyright:
© 2021 Elsevier Ltd

Keywords

Ca signaling
Ca transport
SERCA activation
membrane protein-protein interactions
membrane proteins
oriented samples
solid-state NMR
structural topology

Publisher link

10.1016/j.str.2021.11.003

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title = "A kink in DWORF helical structure controls the activation of the sarcoplasmic reticulum Ca2+-ATPase",

abstract = "SERCA is a P-type ATPase embedded in the sarcoplasmic reticulum and plays a central role in muscle relaxation. SERCA's function is regulated by single-pass membrane proteins called regulins. Unlike other regulins, dwarf open reading frame (DWORF) expressed in cardiac muscle has a unique activating effect. Here, we determine the structure and topology of DWORF in lipid bilayers using a combination of oriented sample solid-state NMR spectroscopy and replica-averaged orientationally restrained molecular dynamics. We found that DWORF's structural topology consists of a dynamic N-terminal domain, an amphipathic juxtamembrane helix that crosses the lipid groups at an angle of 64°, and a transmembrane C-terminal helix with an angle of 32°. A kink induced by Pro15, unique to DWORF, separates the two helical domains. A single Pro15Ala mutant significantly decreases the kink and eliminates DWORF's activating effect on SERCA. Overall, our findings directly link DWORF's structural topology to its activating effect on SERCA.",

keywords = "Ca signaling, Ca transport, SERCA activation, membrane protein-protein interactions, membrane proteins, oriented samples, solid-state NMR, structural topology",

author = "Reddy, \{U. Venkateswara\} and Weber, \{Daniel K\} and Songlin Wang and Erik Larsen and \{De Simone\}, Alfonso and Gopinath Tata and Seth Robia and Gianluigi Veglia",

note = "Funding Information: D.K.W. was supported by an American Heart Association postdoctoral fellowship (19POST34420009). This work was supported by National Heart, Lung, and Blood Institute grants HL143816 to S.R. and G.V. and HL092321 to S.R. We thank Caitlin Walker for her assistance with cloning the DWORF P15A expression construct and Dr. Maximo Sanz-Hern{\'a}ndez for his contributions to the modeling work. Publisher Copyright: {\textcopyright} 2021 Elsevier Ltd",

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T1 - A kink in DWORF helical structure controls the activation of the sarcoplasmic reticulum Ca2+-ATPase

AU - Reddy, U. Venkateswara

AU - Weber, Daniel K

AU - Wang, Songlin

AU - Larsen, Erik

AU - De Simone, Alfonso

AU - Tata, Gopinath

AU - Robia, Seth

AU - Veglia, Gianluigi

N1 - Funding Information: D.K.W. was supported by an American Heart Association postdoctoral fellowship (19POST34420009). This work was supported by National Heart, Lung, and Blood Institute grants HL143816 to S.R. and G.V. and HL092321 to S.R. We thank Caitlin Walker for her assistance with cloning the DWORF P15A expression construct and Dr. Maximo Sanz-Hernández for his contributions to the modeling work. Publisher Copyright: © 2021 Elsevier Ltd

PY - 2022/3/3

Y1 - 2022/3/3

N2 - SERCA is a P-type ATPase embedded in the sarcoplasmic reticulum and plays a central role in muscle relaxation. SERCA's function is regulated by single-pass membrane proteins called regulins. Unlike other regulins, dwarf open reading frame (DWORF) expressed in cardiac muscle has a unique activating effect. Here, we determine the structure and topology of DWORF in lipid bilayers using a combination of oriented sample solid-state NMR spectroscopy and replica-averaged orientationally restrained molecular dynamics. We found that DWORF's structural topology consists of a dynamic N-terminal domain, an amphipathic juxtamembrane helix that crosses the lipid groups at an angle of 64°, and a transmembrane C-terminal helix with an angle of 32°. A kink induced by Pro15, unique to DWORF, separates the two helical domains. A single Pro15Ala mutant significantly decreases the kink and eliminates DWORF's activating effect on SERCA. Overall, our findings directly link DWORF's structural topology to its activating effect on SERCA.

AB - SERCA is a P-type ATPase embedded in the sarcoplasmic reticulum and plays a central role in muscle relaxation. SERCA's function is regulated by single-pass membrane proteins called regulins. Unlike other regulins, dwarf open reading frame (DWORF) expressed in cardiac muscle has a unique activating effect. Here, we determine the structure and topology of DWORF in lipid bilayers using a combination of oriented sample solid-state NMR spectroscopy and replica-averaged orientationally restrained molecular dynamics. We found that DWORF's structural topology consists of a dynamic N-terminal domain, an amphipathic juxtamembrane helix that crosses the lipid groups at an angle of 64°, and a transmembrane C-terminal helix with an angle of 32°. A kink induced by Pro15, unique to DWORF, separates the two helical domains. A single Pro15Ala mutant significantly decreases the kink and eliminates DWORF's activating effect on SERCA. Overall, our findings directly link DWORF's structural topology to its activating effect on SERCA.

KW - Ca signaling

KW - Ca transport

KW - SERCA activation

KW - membrane protein-protein interactions

KW - membrane proteins

KW - oriented samples

KW - solid-state NMR

KW - structural topology

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