Molecular cloning of chick cardiac muscle tensin: Full-length cDNA sequence, expression, and characterization

Su Hao Lo, Qi An, Shideng Bao, Wai Keung Wong, Yuan Liu, Paul A. Janmey, John H. Hartwig, Lan Bo Chen

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Here we describe the molecular cloning of 7.1-kilobase cDNA encoding chick cardiac muscle tensin. It contains an open reading frame of 1,744 amino acid (aa) residues. Sequence analysis reveals that, in addition to the previously noted SH2 domain (Davis, S., Lu, M. L., Lo, S. H., Lin, S., Butler, J. A., Druker, B. J., Roberts, T. M., An, Q., and Chen, L. B. (1991) Science 252, 712-715), tensin contains virtually all of the known sequence (362 aa) of insertin, an actin-capping protein that allows actin monomer to be "inserted" (Schroer, E., and Wegner, A. (1985) Eur. J. Biochem. 153, 515-520). Moreover, tensin shares partial homology with actin (46.7% identity in 30 aa), β-spectrin's actin-binding consensus (40% identity in 26 aa), BCR (40% identity in 25 aa), catenin α (35% identity in 45 aa), synapsin Ia (25.6% identity in 156 aa), IL-3 receptor (20.2% identity in 384 aa), and IL-2/EPO receptors (14% identity in 20 aa). Recombinant full-length tensin, tagged with an influenza-derived epitope, was over-expressed by a baculovirus system and purified to apparent homogeneity. It migrates as a 200-kDa protein in SDS-polyacrylamide gel electrophoresis, similar to the native tensin. The structure of the tensin molecule has been characterized by light scattering, electron microscopy, and gel filtration. Nine monoclonal antibodies recognizing different regions of tensin have been prepared and characterized. The epitope-tagged recombinant tensin gene was subcloned into a pRcCMV vector and transfected into NIH 3T3 cells. Immunofluorescence stainings with monoclonal antibodies specific for chick tensin (not cross-reactive with mouse tensin) showed that the expressed protein is indeed localized at focal contacts, as that of native tensin.

Original languageEnglish (US)
Pages (from-to)22310-22319
Number of pages10
JournalJournal of Biological Chemistry
Volume269
Issue number35
StatePublished - Sep 2 1994
Externally publishedYes

Fingerprint

Cloning
Molecular Cloning
Muscle
Myocardium
Complementary DNA
Amino Acids
Actins
Epitopes
Actin Capping Proteins
Interleukin-3 Receptors
Monoclonal Antibodies
Synapsins
Tensins
Catenins
Spectrin
NIH 3T3 Cells
Electrophoresis
Focal Adhesions
src Homology Domains
Light scattering

ASJC Scopus subject areas

  • Biochemistry

Cite this

Lo, S. H., An, Q., Bao, S., Wong, W. K., Liu, Y., Janmey, P. A., ... Chen, L. B. (1994). Molecular cloning of chick cardiac muscle tensin: Full-length cDNA sequence, expression, and characterization. Journal of Biological Chemistry, 269(35), 22310-22319.

Molecular cloning of chick cardiac muscle tensin : Full-length cDNA sequence, expression, and characterization. / Lo, Su Hao; An, Qi; Bao, Shideng; Wong, Wai Keung; Liu, Yuan; Janmey, Paul A.; Hartwig, John H.; Chen, Lan Bo.

In: Journal of Biological Chemistry, Vol. 269, No. 35, 02.09.1994, p. 22310-22319.

Research output: Contribution to journalArticle

Lo, SH, An, Q, Bao, S, Wong, WK, Liu, Y, Janmey, PA, Hartwig, JH & Chen, LB 1994, 'Molecular cloning of chick cardiac muscle tensin: Full-length cDNA sequence, expression, and characterization', Journal of Biological Chemistry, vol. 269, no. 35, pp. 22310-22319.
Lo, Su Hao ; An, Qi ; Bao, Shideng ; Wong, Wai Keung ; Liu, Yuan ; Janmey, Paul A. ; Hartwig, John H. ; Chen, Lan Bo. / Molecular cloning of chick cardiac muscle tensin : Full-length cDNA sequence, expression, and characterization. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 35. pp. 22310-22319.
@article{73ad5147ef074eaf9408fa0a4d513351,
title = "Molecular cloning of chick cardiac muscle tensin: Full-length cDNA sequence, expression, and characterization",
abstract = "Here we describe the molecular cloning of 7.1-kilobase cDNA encoding chick cardiac muscle tensin. It contains an open reading frame of 1,744 amino acid (aa) residues. Sequence analysis reveals that, in addition to the previously noted SH2 domain (Davis, S., Lu, M. L., Lo, S. H., Lin, S., Butler, J. A., Druker, B. J., Roberts, T. M., An, Q., and Chen, L. B. (1991) Science 252, 712-715), tensin contains virtually all of the known sequence (362 aa) of insertin, an actin-capping protein that allows actin monomer to be {"}inserted{"} (Schroer, E., and Wegner, A. (1985) Eur. J. Biochem. 153, 515-520). Moreover, tensin shares partial homology with actin (46.7{\%} identity in 30 aa), β-spectrin's actin-binding consensus (40{\%} identity in 26 aa), BCR (40{\%} identity in 25 aa), catenin α (35{\%} identity in 45 aa), synapsin Ia (25.6{\%} identity in 156 aa), IL-3 receptor (20.2{\%} identity in 384 aa), and IL-2/EPO receptors (14{\%} identity in 20 aa). Recombinant full-length tensin, tagged with an influenza-derived epitope, was over-expressed by a baculovirus system and purified to apparent homogeneity. It migrates as a 200-kDa protein in SDS-polyacrylamide gel electrophoresis, similar to the native tensin. The structure of the tensin molecule has been characterized by light scattering, electron microscopy, and gel filtration. Nine monoclonal antibodies recognizing different regions of tensin have been prepared and characterized. The epitope-tagged recombinant tensin gene was subcloned into a pRcCMV vector and transfected into NIH 3T3 cells. Immunofluorescence stainings with monoclonal antibodies specific for chick tensin (not cross-reactive with mouse tensin) showed that the expressed protein is indeed localized at focal contacts, as that of native tensin.",
author = "Lo, {Su Hao} and Qi An and Shideng Bao and Wong, {Wai Keung} and Yuan Liu and Janmey, {Paul A.} and Hartwig, {John H.} and Chen, {Lan Bo}",
year = "1994",
month = "9",
day = "2",
language = "English (US)",
volume = "269",
pages = "22310--22319",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "35",

}

TY - JOUR

T1 - Molecular cloning of chick cardiac muscle tensin

T2 - Full-length cDNA sequence, expression, and characterization

AU - Lo, Su Hao

AU - An, Qi

AU - Bao, Shideng

AU - Wong, Wai Keung

AU - Liu, Yuan

AU - Janmey, Paul A.

AU - Hartwig, John H.

AU - Chen, Lan Bo

PY - 1994/9/2

Y1 - 1994/9/2

N2 - Here we describe the molecular cloning of 7.1-kilobase cDNA encoding chick cardiac muscle tensin. It contains an open reading frame of 1,744 amino acid (aa) residues. Sequence analysis reveals that, in addition to the previously noted SH2 domain (Davis, S., Lu, M. L., Lo, S. H., Lin, S., Butler, J. A., Druker, B. J., Roberts, T. M., An, Q., and Chen, L. B. (1991) Science 252, 712-715), tensin contains virtually all of the known sequence (362 aa) of insertin, an actin-capping protein that allows actin monomer to be "inserted" (Schroer, E., and Wegner, A. (1985) Eur. J. Biochem. 153, 515-520). Moreover, tensin shares partial homology with actin (46.7% identity in 30 aa), β-spectrin's actin-binding consensus (40% identity in 26 aa), BCR (40% identity in 25 aa), catenin α (35% identity in 45 aa), synapsin Ia (25.6% identity in 156 aa), IL-3 receptor (20.2% identity in 384 aa), and IL-2/EPO receptors (14% identity in 20 aa). Recombinant full-length tensin, tagged with an influenza-derived epitope, was over-expressed by a baculovirus system and purified to apparent homogeneity. It migrates as a 200-kDa protein in SDS-polyacrylamide gel electrophoresis, similar to the native tensin. The structure of the tensin molecule has been characterized by light scattering, electron microscopy, and gel filtration. Nine monoclonal antibodies recognizing different regions of tensin have been prepared and characterized. The epitope-tagged recombinant tensin gene was subcloned into a pRcCMV vector and transfected into NIH 3T3 cells. Immunofluorescence stainings with monoclonal antibodies specific for chick tensin (not cross-reactive with mouse tensin) showed that the expressed protein is indeed localized at focal contacts, as that of native tensin.

AB - Here we describe the molecular cloning of 7.1-kilobase cDNA encoding chick cardiac muscle tensin. It contains an open reading frame of 1,744 amino acid (aa) residues. Sequence analysis reveals that, in addition to the previously noted SH2 domain (Davis, S., Lu, M. L., Lo, S. H., Lin, S., Butler, J. A., Druker, B. J., Roberts, T. M., An, Q., and Chen, L. B. (1991) Science 252, 712-715), tensin contains virtually all of the known sequence (362 aa) of insertin, an actin-capping protein that allows actin monomer to be "inserted" (Schroer, E., and Wegner, A. (1985) Eur. J. Biochem. 153, 515-520). Moreover, tensin shares partial homology with actin (46.7% identity in 30 aa), β-spectrin's actin-binding consensus (40% identity in 26 aa), BCR (40% identity in 25 aa), catenin α (35% identity in 45 aa), synapsin Ia (25.6% identity in 156 aa), IL-3 receptor (20.2% identity in 384 aa), and IL-2/EPO receptors (14% identity in 20 aa). Recombinant full-length tensin, tagged with an influenza-derived epitope, was over-expressed by a baculovirus system and purified to apparent homogeneity. It migrates as a 200-kDa protein in SDS-polyacrylamide gel electrophoresis, similar to the native tensin. The structure of the tensin molecule has been characterized by light scattering, electron microscopy, and gel filtration. Nine monoclonal antibodies recognizing different regions of tensin have been prepared and characterized. The epitope-tagged recombinant tensin gene was subcloned into a pRcCMV vector and transfected into NIH 3T3 cells. Immunofluorescence stainings with monoclonal antibodies specific for chick tensin (not cross-reactive with mouse tensin) showed that the expressed protein is indeed localized at focal contacts, as that of native tensin.

UR - http://www.scopus.com/inward/record.url?scp=0027931203&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027931203&partnerID=8YFLogxK

M3 - Article

C2 - 8071358

AN - SCOPUS:0027931203

VL - 269

SP - 22310

EP - 22319

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 35

ER -