Sarcoplasmic reticulum from horse gluteal muscle is poised for enhanced calcium transport

Joseph M. Autry; Bengt Svensson; Samuel F. Carlson; Zhenhui Chen; Razvan L. Cornea; David D. Thomas; Stephanie J. Valberg

doi:10.3390/vetsci8120289

Sarcoplasmic reticulum from horse gluteal muscle is poised for enhanced calcium transport

Joseph M. Autry, Bengt Svensson, Samuel F. Carlson, Zhenhui Chen, Razvan L. Cornea, David D. Thomas, Stephanie J. Valberg

Chemical and Structural Biology (TMED)

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1 Scopus citations

Abstract

We have analyzed the enzymatic activity of the sarcoplasmic reticulum (SR) Ca²⁺-transporting ATPase (SERCA) from the horse gluteal muscle. Horses are bred for peak athletic performance yet exhibit a high incidence of exertional rhabdomyolysis, with elevated levels of cytosolic Ca²⁺ proposed as a correlative linkage. We recently reported an improved protocol for isolating SR vesicles from horse muscle; these horse SR vesicles contain an abundant level of SERCA and only trace-levels of sarcolipin (SLN), the inhibitory peptide subunit of SERCA in mammalian fast-twitch skeletal muscle. Here, we report that the in vitro Ca²⁺ transport rate of horse SR vesicles is 2.3 ± 0.7-fold greater than rabbit SR vesicles, which express close to equimolar levels of SERCA and SLN. This suggests that horse myofibers exhibit an enhanced SR Ca²⁺ transport rate and increased luminal Ca²⁺ stores in vivo. Using the densitometry of Coomassie-stained SDS-PAGE gels, we determined that horse SR vesicles express an abundant level of the luminal SR Ca²⁺ storage protein calsequestrin (CASQ), with a CASQ-to-SERCA ratio about double that in rabbit SR vesicles. Thus, we propose that SR Ca²⁺ cycling in horse myofibers is enhanced by a reduced SLN inhibition of SERCA and by an abundant expression of CASQ. Together, these results suggest that horse muscle contractility and susceptibility to exertional rhabdomyolysis are promoted by enhanced SR Ca²⁺ uptake and luminal Ca²⁺ storage.

Original language	English (US)
Article number	289
Journal	Veterinary Sciences
Volume	8
Issue number	12
DOIs	https://doi.org/10.3390/vetsci8120289
State	Published - Dec 2021

Bibliographical note

Funding Information:
Funding: This study was supported in part by a Morris Animal Foundation grant to S.J.V, J.M.A., and D.D.T. (grant number D16EQ-004). Morris Animal Foundation is the global leader in supporting science that advances animal health. This study was supported in part by National Institutes of Health grants to D.D.T. (grant numbers GM027906 and AG026160) and to D.D.T. and R.L.C. (HL139065). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. This study was supported in part by an American Heart Association grant to Z.C. (grant number 18TPA34170284/Chen, Zhenhui/2018). The funding agencies had no role in study design, data collection, data analysis, manuscript preparation, or decision to publish.

Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Keywords

Calcium regulation
Calsequestrin
Comparative biochemistry
Excitation–contraction coupling
Exertional rhabdomyolysis
Intraluminal protein
Ion-motive ATPase
Membrane vesicles
Regulatory subunit
Skeletal muscle

PubMed: MeSH publication types

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10.3390/vetsci8120289

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@article{227130a9a5e640aabec0d92bef3e06c0,

title = "Sarcoplasmic reticulum from horse gluteal muscle is poised for enhanced calcium transport",

abstract = "We have analyzed the enzymatic activity of the sarcoplasmic reticulum (SR) Ca2+-transporting ATPase (SERCA) from the horse gluteal muscle. Horses are bred for peak athletic performance yet exhibit a high incidence of exertional rhabdomyolysis, with elevated levels of cytosolic Ca2+ proposed as a correlative linkage. We recently reported an improved protocol for isolating SR vesicles from horse muscle; these horse SR vesicles contain an abundant level of SERCA and only trace-levels of sarcolipin (SLN), the inhibitory peptide subunit of SERCA in mammalian fast-twitch skeletal muscle. Here, we report that the in vitro Ca2+ transport rate of horse SR vesicles is 2.3 ± 0.7-fold greater than rabbit SR vesicles, which express close to equimolar levels of SERCA and SLN. This suggests that horse myofibers exhibit an enhanced SR Ca2+ transport rate and increased luminal Ca2+ stores in vivo. Using the densitometry of Coomassie-stained SDS-PAGE gels, we determined that horse SR vesicles express an abundant level of the luminal SR Ca2+ storage protein calsequestrin (CASQ), with a CASQ-to-SERCA ratio about double that in rabbit SR vesicles. Thus, we propose that SR Ca2+ cycling in horse myofibers is enhanced by a reduced SLN inhibition of SERCA and by an abundant expression of CASQ. Together, these results suggest that horse muscle contractility and susceptibility to exertional rhabdomyolysis are promoted by enhanced SR Ca2+ uptake and luminal Ca2+ storage.",

keywords = "Calcium regulation, Calsequestrin, Comparative biochemistry, Excitation–contraction coupling, Exertional rhabdomyolysis, Intraluminal protein, Ion-motive ATPase, Membrane vesicles, Regulatory subunit, Skeletal muscle",

author = "Autry, {Joseph M.} and Bengt Svensson and Carlson, {Samuel F.} and Zhenhui Chen and Cornea, {Razvan L.} and Thomas, {David D.} and Valberg, {Stephanie J.}",

note = "Funding Information: Funding: This study was supported in part by a Morris Animal Foundation grant to S.J.V, J.M.A., and D.D.T. (grant number D16EQ-004). Morris Animal Foundation is the global leader in supporting science that advances animal health. This study was supported in part by National Institutes of Health grants to D.D.T. (grant numbers GM027906 and AG026160) and to D.D.T. and R.L.C. (HL139065). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. This study was supported in part by an American Heart Association grant to Z.C. (grant number 18TPA34170284/Chen, Zhenhui/2018). The funding agencies had no role in study design, data collection, data analysis, manuscript preparation, or decision to publish. Publisher Copyright: {\textcopyright} 2021 by the authors. Licensee MDPI, Basel, Switzerland.",

year = "2021",

month = dec,

doi = "10.3390/vetsci8120289",

language = "English (US)",

volume = "8",

journal = "Veterinary Sciences",

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T1 - Sarcoplasmic reticulum from horse gluteal muscle is poised for enhanced calcium transport

AU - Autry, Joseph M.

AU - Svensson, Bengt

AU - Carlson, Samuel F.

AU - Chen, Zhenhui

AU - Cornea, Razvan L.

AU - Thomas, David D.

AU - Valberg, Stephanie J.

N1 - Funding Information: Funding: This study was supported in part by a Morris Animal Foundation grant to S.J.V, J.M.A., and D.D.T. (grant number D16EQ-004). Morris Animal Foundation is the global leader in supporting science that advances animal health. This study was supported in part by National Institutes of Health grants to D.D.T. (grant numbers GM027906 and AG026160) and to D.D.T. and R.L.C. (HL139065). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. This study was supported in part by an American Heart Association grant to Z.C. (grant number 18TPA34170284/Chen, Zhenhui/2018). The funding agencies had no role in study design, data collection, data analysis, manuscript preparation, or decision to publish. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

PY - 2021/12

Y1 - 2021/12

N2 - We have analyzed the enzymatic activity of the sarcoplasmic reticulum (SR) Ca2+-transporting ATPase (SERCA) from the horse gluteal muscle. Horses are bred for peak athletic performance yet exhibit a high incidence of exertional rhabdomyolysis, with elevated levels of cytosolic Ca2+ proposed as a correlative linkage. We recently reported an improved protocol for isolating SR vesicles from horse muscle; these horse SR vesicles contain an abundant level of SERCA and only trace-levels of sarcolipin (SLN), the inhibitory peptide subunit of SERCA in mammalian fast-twitch skeletal muscle. Here, we report that the in vitro Ca2+ transport rate of horse SR vesicles is 2.3 ± 0.7-fold greater than rabbit SR vesicles, which express close to equimolar levels of SERCA and SLN. This suggests that horse myofibers exhibit an enhanced SR Ca2+ transport rate and increased luminal Ca2+ stores in vivo. Using the densitometry of Coomassie-stained SDS-PAGE gels, we determined that horse SR vesicles express an abundant level of the luminal SR Ca2+ storage protein calsequestrin (CASQ), with a CASQ-to-SERCA ratio about double that in rabbit SR vesicles. Thus, we propose that SR Ca2+ cycling in horse myofibers is enhanced by a reduced SLN inhibition of SERCA and by an abundant expression of CASQ. Together, these results suggest that horse muscle contractility and susceptibility to exertional rhabdomyolysis are promoted by enhanced SR Ca2+ uptake and luminal Ca2+ storage.

AB - We have analyzed the enzymatic activity of the sarcoplasmic reticulum (SR) Ca2+-transporting ATPase (SERCA) from the horse gluteal muscle. Horses are bred for peak athletic performance yet exhibit a high incidence of exertional rhabdomyolysis, with elevated levels of cytosolic Ca2+ proposed as a correlative linkage. We recently reported an improved protocol for isolating SR vesicles from horse muscle; these horse SR vesicles contain an abundant level of SERCA and only trace-levels of sarcolipin (SLN), the inhibitory peptide subunit of SERCA in mammalian fast-twitch skeletal muscle. Here, we report that the in vitro Ca2+ transport rate of horse SR vesicles is 2.3 ± 0.7-fold greater than rabbit SR vesicles, which express close to equimolar levels of SERCA and SLN. This suggests that horse myofibers exhibit an enhanced SR Ca2+ transport rate and increased luminal Ca2+ stores in vivo. Using the densitometry of Coomassie-stained SDS-PAGE gels, we determined that horse SR vesicles express an abundant level of the luminal SR Ca2+ storage protein calsequestrin (CASQ), with a CASQ-to-SERCA ratio about double that in rabbit SR vesicles. Thus, we propose that SR Ca2+ cycling in horse myofibers is enhanced by a reduced SLN inhibition of SERCA and by an abundant expression of CASQ. Together, these results suggest that horse muscle contractility and susceptibility to exertional rhabdomyolysis are promoted by enhanced SR Ca2+ uptake and luminal Ca2+ storage.

KW - Calcium regulation

KW - Calsequestrin

KW - Comparative biochemistry

KW - Excitation–contraction coupling

KW - Exertional rhabdomyolysis

KW - Intraluminal protein

KW - Ion-motive ATPase

KW - Membrane vesicles

KW - Regulatory subunit

KW - Skeletal muscle

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U2 - 10.3390/vetsci8120289

DO - 10.3390/vetsci8120289

M3 - Article

C2 - 34941816

AN - SCOPUS:85119983666

SN - 2306-7381

VL - 8

JO - Veterinary Sciences

JF - Veterinary Sciences

IS - 12

M1 - 289

ER -

Sarcoplasmic reticulum from horse gluteal muscle is poised for enhanced calcium transport

Abstract

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Keywords

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