Biochemical characterization and molecular cloning of cardiac triadin

Wei Guo, Annelise O. Jorgensen, Larry Jones, Kevin P. Campbell

Research output: Contribution to journalArticle

85 Citations (Scopus)

Abstract

Triadin is an intrinsic membrane protein first identified in the skeletal muscle junctional sarcoplasmic reticulum and is considered to play an important role in excitation-contraction coupling. Using polyclonal antibodies to skeletal muscle triadin, we have identified and characterized three isoforms in rabbit cardiac muscle. The cDNAs encoding these three isoforms of triadin have been isolated by reverse transcription-polymerase chain reaction and cDNA library screening. The deduced amino acid sequences show that these proteins are identical in their N-terminal sequences, whereas the C-terminal sequences are distinct from each other and from that of skeletal muscle triadin. Based upon both the amino acid sequences and biochemical analysis, all three triadin isoforms share similar membrane topology with skeletal muscle triadin. Immunofluorescence staining of rabbit cardiac muscle with antibodies purified from the homologous region of triadin shows that cardiac triadin is primarily confined to the I-band region of cardiac myocytes, where the junctional and corbular sarcoplasmic reticulum is located. Furthermore, we demonstrate that the conserved region of the luminal domain of triadin is able to bind both the ryanodine receptor and calsequestrin in cardiac muscle. These results suggest that triadin colocalizes with and binds to the ryanodine receptor and calsequestrin and carries out a function in the lumen of the junctional sarcoplasmic reticulum that is important for both skeletal and cardiac muscle excitation- contraction coupling.

Original languageEnglish
Pages (from-to)458-465
Number of pages8
JournalJournal of Biological Chemistry
Volume271
Issue number1
DOIs
StatePublished - Jan 5 1996

Fingerprint

Cloning
Molecular Cloning
Muscle
Skeletal Muscle
Myocardium
Sarcoplasmic Reticulum
Calsequestrin
Excitation Contraction Coupling
Protein Isoforms
Ryanodine Receptor Calcium Release Channel
triadin
Rabbits
Amino Acids
Antibodies
Polymerase chain reaction
Protein Sequence Analysis
Transcription
Muscle Contraction
Gene Library
Cardiac Myocytes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Biochemical characterization and molecular cloning of cardiac triadin. / Guo, Wei; Jorgensen, Annelise O.; Jones, Larry; Campbell, Kevin P.

In: Journal of Biological Chemistry, Vol. 271, No. 1, 05.01.1996, p. 458-465.

Research output: Contribution to journalArticle

Guo, Wei ; Jorgensen, Annelise O. ; Jones, Larry ; Campbell, Kevin P. / Biochemical characterization and molecular cloning of cardiac triadin. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 1. pp. 458-465.
@article{0658923a194c4f2fb7d351163e678970,
title = "Biochemical characterization and molecular cloning of cardiac triadin",
abstract = "Triadin is an intrinsic membrane protein first identified in the skeletal muscle junctional sarcoplasmic reticulum and is considered to play an important role in excitation-contraction coupling. Using polyclonal antibodies to skeletal muscle triadin, we have identified and characterized three isoforms in rabbit cardiac muscle. The cDNAs encoding these three isoforms of triadin have been isolated by reverse transcription-polymerase chain reaction and cDNA library screening. The deduced amino acid sequences show that these proteins are identical in their N-terminal sequences, whereas the C-terminal sequences are distinct from each other and from that of skeletal muscle triadin. Based upon both the amino acid sequences and biochemical analysis, all three triadin isoforms share similar membrane topology with skeletal muscle triadin. Immunofluorescence staining of rabbit cardiac muscle with antibodies purified from the homologous region of triadin shows that cardiac triadin is primarily confined to the I-band region of cardiac myocytes, where the junctional and corbular sarcoplasmic reticulum is located. Furthermore, we demonstrate that the conserved region of the luminal domain of triadin is able to bind both the ryanodine receptor and calsequestrin in cardiac muscle. These results suggest that triadin colocalizes with and binds to the ryanodine receptor and calsequestrin and carries out a function in the lumen of the junctional sarcoplasmic reticulum that is important for both skeletal and cardiac muscle excitation- contraction coupling.",
author = "Wei Guo and Jorgensen, {Annelise O.} and Larry Jones and Campbell, {Kevin P.}",
year = "1996",
month = "1",
day = "5",
doi = "10.1074/jbc.271.1.458",
language = "English",
volume = "271",
pages = "458--465",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "1",

}

TY - JOUR

T1 - Biochemical characterization and molecular cloning of cardiac triadin

AU - Guo, Wei

AU - Jorgensen, Annelise O.

AU - Jones, Larry

AU - Campbell, Kevin P.

PY - 1996/1/5

Y1 - 1996/1/5

N2 - Triadin is an intrinsic membrane protein first identified in the skeletal muscle junctional sarcoplasmic reticulum and is considered to play an important role in excitation-contraction coupling. Using polyclonal antibodies to skeletal muscle triadin, we have identified and characterized three isoforms in rabbit cardiac muscle. The cDNAs encoding these three isoforms of triadin have been isolated by reverse transcription-polymerase chain reaction and cDNA library screening. The deduced amino acid sequences show that these proteins are identical in their N-terminal sequences, whereas the C-terminal sequences are distinct from each other and from that of skeletal muscle triadin. Based upon both the amino acid sequences and biochemical analysis, all three triadin isoforms share similar membrane topology with skeletal muscle triadin. Immunofluorescence staining of rabbit cardiac muscle with antibodies purified from the homologous region of triadin shows that cardiac triadin is primarily confined to the I-band region of cardiac myocytes, where the junctional and corbular sarcoplasmic reticulum is located. Furthermore, we demonstrate that the conserved region of the luminal domain of triadin is able to bind both the ryanodine receptor and calsequestrin in cardiac muscle. These results suggest that triadin colocalizes with and binds to the ryanodine receptor and calsequestrin and carries out a function in the lumen of the junctional sarcoplasmic reticulum that is important for both skeletal and cardiac muscle excitation- contraction coupling.

AB - Triadin is an intrinsic membrane protein first identified in the skeletal muscle junctional sarcoplasmic reticulum and is considered to play an important role in excitation-contraction coupling. Using polyclonal antibodies to skeletal muscle triadin, we have identified and characterized three isoforms in rabbit cardiac muscle. The cDNAs encoding these three isoforms of triadin have been isolated by reverse transcription-polymerase chain reaction and cDNA library screening. The deduced amino acid sequences show that these proteins are identical in their N-terminal sequences, whereas the C-terminal sequences are distinct from each other and from that of skeletal muscle triadin. Based upon both the amino acid sequences and biochemical analysis, all three triadin isoforms share similar membrane topology with skeletal muscle triadin. Immunofluorescence staining of rabbit cardiac muscle with antibodies purified from the homologous region of triadin shows that cardiac triadin is primarily confined to the I-band region of cardiac myocytes, where the junctional and corbular sarcoplasmic reticulum is located. Furthermore, we demonstrate that the conserved region of the luminal domain of triadin is able to bind both the ryanodine receptor and calsequestrin in cardiac muscle. These results suggest that triadin colocalizes with and binds to the ryanodine receptor and calsequestrin and carries out a function in the lumen of the junctional sarcoplasmic reticulum that is important for both skeletal and cardiac muscle excitation- contraction coupling.

UR - http://www.scopus.com/inward/record.url?scp=0030039603&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030039603&partnerID=8YFLogxK

U2 - 10.1074/jbc.271.1.458

DO - 10.1074/jbc.271.1.458

M3 - Article

C2 - 8550602

AN - SCOPUS:0030039603

VL - 271

SP - 458

EP - 465

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 1

ER -