Association of small ankyrin 1 with the sarcoplasmic reticulum

Neil C. Porter, Wendy G. Resneck, Andrea O'Neill, Damian B. Van Rossum, Michele R. Stone, Robert J. Bloch

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Small ankyrin 1, or sAnk1, is a small, alternatively spliced product of the erythroid ankyrin gene, ANK1, that is expressed in striated muscle and concentrated in the network sarcoplasmic reticulum (SR) surrounding the Z disks and M lines. We have characterized sAnk1 in muscle homogenates and SR vesicles, and have identified the region that targets it to the network SR. Selective extractions and partitioning into Triton X-114 show that sAnk1 behaves like the SR Ca-ATPase and so is an integral protein of the SR membrane. Mild proteolytic treatment of isolated SR vesicles indicates that sAnk1 is oriented with its hydrophilic, C-terminal sequence exposed to the solution, which is equivalent to the cytoplasmic face of the SR membrane in situ. SDS-PAGE in non-reducing gels suggests that sAnk1 is present as dimers and larger oligomers in the native SR. These results suggest that sAnk1 is oligomeric and oriented with its C-terminus exposed to the cytoplasm, where it may interact with proteins of the contractile apparatus. The N-terminal 29 amino acid hydrophobic sequence of sAnk1, which is predicted to span the SR membrane, is sufficient to target proteins to and anchor them in internal membranes of HEK 293 cells. It also targets reporter proteins to the network SR of skeletal myofibers and is thus the first example of a sequence that targets proteins to a particular compartment of the SR.

Original languageEnglish (US)
Pages (from-to)421-432
Number of pages12
JournalMolecular Membrane Biology
Volume22
Issue number5
DOIs
StatePublished - Sep 1 2005

Fingerprint

Ankyrins
Sarcoplasmic Reticulum
Membranes
Proteins
Contractile Proteins
Striated Muscle
HEK293 Cells
Adenosine Triphosphatases
Polyacrylamide Gel Electrophoresis
Amino Acid Sequence
Cytoplasm

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Porter, N. C., Resneck, W. G., O'Neill, A., Van Rossum, D. B., Stone, M. R., & Bloch, R. J. (2005). Association of small ankyrin 1 with the sarcoplasmic reticulum. Molecular Membrane Biology, 22(5), 421-432. https://doi.org/10.1080/09687860500244262
Porter, Neil C. ; Resneck, Wendy G. ; O'Neill, Andrea ; Van Rossum, Damian B. ; Stone, Michele R. ; Bloch, Robert J. / Association of small ankyrin 1 with the sarcoplasmic reticulum. In: Molecular Membrane Biology. 2005 ; Vol. 22, No. 5. pp. 421-432.
@article{960b91e75457406d84697ae161decfe5,
title = "Association of small ankyrin 1 with the sarcoplasmic reticulum",
abstract = "Small ankyrin 1, or sAnk1, is a small, alternatively spliced product of the erythroid ankyrin gene, ANK1, that is expressed in striated muscle and concentrated in the network sarcoplasmic reticulum (SR) surrounding the Z disks and M lines. We have characterized sAnk1 in muscle homogenates and SR vesicles, and have identified the region that targets it to the network SR. Selective extractions and partitioning into Triton X-114 show that sAnk1 behaves like the SR Ca-ATPase and so is an integral protein of the SR membrane. Mild proteolytic treatment of isolated SR vesicles indicates that sAnk1 is oriented with its hydrophilic, C-terminal sequence exposed to the solution, which is equivalent to the cytoplasmic face of the SR membrane in situ. SDS-PAGE in non-reducing gels suggests that sAnk1 is present as dimers and larger oligomers in the native SR. These results suggest that sAnk1 is oligomeric and oriented with its C-terminus exposed to the cytoplasm, where it may interact with proteins of the contractile apparatus. The N-terminal 29 amino acid hydrophobic sequence of sAnk1, which is predicted to span the SR membrane, is sufficient to target proteins to and anchor them in internal membranes of HEK 293 cells. It also targets reporter proteins to the network SR of skeletal myofibers and is thus the first example of a sequence that targets proteins to a particular compartment of the SR.",
author = "Porter, {Neil C.} and Resneck, {Wendy G.} and Andrea O'Neill and {Van Rossum}, {Damian B.} and Stone, {Michele R.} and Bloch, {Robert J.}",
year = "2005",
month = "9",
day = "1",
doi = "10.1080/09687860500244262",
language = "English (US)",
volume = "22",
pages = "421--432",
journal = "Molecular Membrane Biology",
issn = "0968-7688",
publisher = "Informa Healthcare",
number = "5",

}

Porter, NC, Resneck, WG, O'Neill, A, Van Rossum, DB, Stone, MR & Bloch, RJ 2005, 'Association of small ankyrin 1 with the sarcoplasmic reticulum', Molecular Membrane Biology, vol. 22, no. 5, pp. 421-432. https://doi.org/10.1080/09687860500244262

Association of small ankyrin 1 with the sarcoplasmic reticulum. / Porter, Neil C.; Resneck, Wendy G.; O'Neill, Andrea; Van Rossum, Damian B.; Stone, Michele R.; Bloch, Robert J.

In: Molecular Membrane Biology, Vol. 22, No. 5, 01.09.2005, p. 421-432.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Association of small ankyrin 1 with the sarcoplasmic reticulum

AU - Porter, Neil C.

AU - Resneck, Wendy G.

AU - O'Neill, Andrea

AU - Van Rossum, Damian B.

AU - Stone, Michele R.

AU - Bloch, Robert J.

PY - 2005/9/1

Y1 - 2005/9/1

N2 - Small ankyrin 1, or sAnk1, is a small, alternatively spliced product of the erythroid ankyrin gene, ANK1, that is expressed in striated muscle and concentrated in the network sarcoplasmic reticulum (SR) surrounding the Z disks and M lines. We have characterized sAnk1 in muscle homogenates and SR vesicles, and have identified the region that targets it to the network SR. Selective extractions and partitioning into Triton X-114 show that sAnk1 behaves like the SR Ca-ATPase and so is an integral protein of the SR membrane. Mild proteolytic treatment of isolated SR vesicles indicates that sAnk1 is oriented with its hydrophilic, C-terminal sequence exposed to the solution, which is equivalent to the cytoplasmic face of the SR membrane in situ. SDS-PAGE in non-reducing gels suggests that sAnk1 is present as dimers and larger oligomers in the native SR. These results suggest that sAnk1 is oligomeric and oriented with its C-terminus exposed to the cytoplasm, where it may interact with proteins of the contractile apparatus. The N-terminal 29 amino acid hydrophobic sequence of sAnk1, which is predicted to span the SR membrane, is sufficient to target proteins to and anchor them in internal membranes of HEK 293 cells. It also targets reporter proteins to the network SR of skeletal myofibers and is thus the first example of a sequence that targets proteins to a particular compartment of the SR.

AB - Small ankyrin 1, or sAnk1, is a small, alternatively spliced product of the erythroid ankyrin gene, ANK1, that is expressed in striated muscle and concentrated in the network sarcoplasmic reticulum (SR) surrounding the Z disks and M lines. We have characterized sAnk1 in muscle homogenates and SR vesicles, and have identified the region that targets it to the network SR. Selective extractions and partitioning into Triton X-114 show that sAnk1 behaves like the SR Ca-ATPase and so is an integral protein of the SR membrane. Mild proteolytic treatment of isolated SR vesicles indicates that sAnk1 is oriented with its hydrophilic, C-terminal sequence exposed to the solution, which is equivalent to the cytoplasmic face of the SR membrane in situ. SDS-PAGE in non-reducing gels suggests that sAnk1 is present as dimers and larger oligomers in the native SR. These results suggest that sAnk1 is oligomeric and oriented with its C-terminus exposed to the cytoplasm, where it may interact with proteins of the contractile apparatus. The N-terminal 29 amino acid hydrophobic sequence of sAnk1, which is predicted to span the SR membrane, is sufficient to target proteins to and anchor them in internal membranes of HEK 293 cells. It also targets reporter proteins to the network SR of skeletal myofibers and is thus the first example of a sequence that targets proteins to a particular compartment of the SR.

UR - http://www.scopus.com/inward/record.url?scp=28544435631&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=28544435631&partnerID=8YFLogxK

U2 - 10.1080/09687860500244262

DO - 10.1080/09687860500244262

M3 - Article

C2 - 16308276

AN - SCOPUS:28544435631

VL - 22

SP - 421

EP - 432

JO - Molecular Membrane Biology

JF - Molecular Membrane Biology

SN - 0968-7688

IS - 5

ER -