Nuclear proteins of the bovine esophageal epithelium: I. Monoclonal antibody W2 specifically reacts with condensed nuclei of differentiated superficial cells

Tang K. Tang, Tse Ming Hong, Chen Yong Lin, Mei Ling Lai, Catherine H L Liu, Hsiu Jong Lo, Ming Eng Wang, Lan Bo Chen, Wen Tien Chen, Wallace Ip, Diane C. Lin, Jim J C Lin, Shin Lin, Tung Tien Sun, Eugenia Wang, John L. Wang, Reen Wu, Cheng Wen Wu, Shu Chien

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Cells from three layers of the bovine esophageal epithelium, representing different stages of differentiation, were dissociated and separated by Percoll gradient centrifugation into fractions of small, medium and large sizes. A majority of the large cells possessed condensed nuclei, a characteristic feature of terminal differentiation of the superficial epithelium. The small cells resembled the proliferate cells of the basal layer. In vitro culture of the esophageal epithelial cells resulted in proliferation of the small cells, colony formation, and, in some cases, differentiation into cells with condensed nuclei. Nuclei, or nuclear subfractions derived from cells of the different layers, were used as immunogens for the generation of hybridomas secreting monoclonal antibodies that bound specifically to different regions of the esophageal tissue. One such antibody, designated W2, labeled the condensed nuclei from the superficial layer of stratified esophageal and corneal epithelia in situ, as well as the large cells from esophageal culture in vitro. Thus, the expression of the W2 antigen may be associated with the process of nuclear condensation during epithelial differentiation. Immunoisolation of the target antigen of W2 from extracts of large cells of the bovine esophagus yielded a band of Mr ∼33,000 on nonreducing polyacrylamide gels. This band dissociated into two polypeptides, of Mr ∼22,000 and ∼11,000, upon treatment with dithiothreitol. Amino acid sequence analysis of the larger polypeptide showed extensive homology to a group of small calcium-binding proteins, including two helix-turn-helix motifs designated as the EF-hand, characteristic of the configuration of the metal-ion coordinating ligands of the calcium-binding site. Similarly, the sequence at the amino terminus of the polypeptide of ∼11,000 indicated that it was the light chain counterpart of the same calcium-binding protein complex.

Original languageEnglish (US)
Pages (from-to)237-247
Number of pages11
JournalJournal of Cell Science
Volume104
Issue number2
StatePublished - Feb 1993
Externally publishedYes

Fingerprint

Nuclear Proteins
Epithelium
Monoclonal Antibodies
Calcium-Binding Proteins
Peptides
Helix-Turn-Helix Motifs
EF Hand Motifs
Antigens
Corneal Epithelium
Dithiothreitol
Protein Sequence Analysis
Hybridomas
Cell Extracts
Centrifugation
Esophagus
Cell Differentiation
Cell Culture Techniques
Epithelial Cells
Metals
Binding Sites

Keywords

  • Calcium binding protein
  • Condensed nuclei
  • Epithelial cell differentiation

ASJC Scopus subject areas

  • Cell Biology

Cite this

Nuclear proteins of the bovine esophageal epithelium : I. Monoclonal antibody W2 specifically reacts with condensed nuclei of differentiated superficial cells. / Tang, Tang K.; Hong, Tse Ming; Lin, Chen Yong; Lai, Mei Ling; Liu, Catherine H L; Lo, Hsiu Jong; Wang, Ming Eng; Chen, Lan Bo; Chen, Wen Tien; Ip, Wallace; Lin, Diane C.; Lin, Jim J C; Lin, Shin; Sun, Tung Tien; Wang, Eugenia; Wang, John L.; Wu, Reen; Wu, Cheng Wen; Chien, Shu.

In: Journal of Cell Science, Vol. 104, No. 2, 02.1993, p. 237-247.

Research output: Contribution to journalArticle

Tang, TK, Hong, TM, Lin, CY, Lai, ML, Liu, CHL, Lo, HJ, Wang, ME, Chen, LB, Chen, WT, Ip, W, Lin, DC, Lin, JJC, Lin, S, Sun, TT, Wang, E, Wang, JL, Wu, R, Wu, CW & Chien, S 1993, 'Nuclear proteins of the bovine esophageal epithelium: I. Monoclonal antibody W2 specifically reacts with condensed nuclei of differentiated superficial cells', Journal of Cell Science, vol. 104, no. 2, pp. 237-247.
Tang, Tang K. ; Hong, Tse Ming ; Lin, Chen Yong ; Lai, Mei Ling ; Liu, Catherine H L ; Lo, Hsiu Jong ; Wang, Ming Eng ; Chen, Lan Bo ; Chen, Wen Tien ; Ip, Wallace ; Lin, Diane C. ; Lin, Jim J C ; Lin, Shin ; Sun, Tung Tien ; Wang, Eugenia ; Wang, John L. ; Wu, Reen ; Wu, Cheng Wen ; Chien, Shu. / Nuclear proteins of the bovine esophageal epithelium : I. Monoclonal antibody W2 specifically reacts with condensed nuclei of differentiated superficial cells. In: Journal of Cell Science. 1993 ; Vol. 104, No. 2. pp. 237-247.
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abstract = "Cells from three layers of the bovine esophageal epithelium, representing different stages of differentiation, were dissociated and separated by Percoll gradient centrifugation into fractions of small, medium and large sizes. A majority of the large cells possessed condensed nuclei, a characteristic feature of terminal differentiation of the superficial epithelium. The small cells resembled the proliferate cells of the basal layer. In vitro culture of the esophageal epithelial cells resulted in proliferation of the small cells, colony formation, and, in some cases, differentiation into cells with condensed nuclei. Nuclei, or nuclear subfractions derived from cells of the different layers, were used as immunogens for the generation of hybridomas secreting monoclonal antibodies that bound specifically to different regions of the esophageal tissue. One such antibody, designated W2, labeled the condensed nuclei from the superficial layer of stratified esophageal and corneal epithelia in situ, as well as the large cells from esophageal culture in vitro. Thus, the expression of the W2 antigen may be associated with the process of nuclear condensation during epithelial differentiation. Immunoisolation of the target antigen of W2 from extracts of large cells of the bovine esophagus yielded a band of Mr ∼33,000 on nonreducing polyacrylamide gels. This band dissociated into two polypeptides, of Mr ∼22,000 and ∼11,000, upon treatment with dithiothreitol. Amino acid sequence analysis of the larger polypeptide showed extensive homology to a group of small calcium-binding proteins, including two helix-turn-helix motifs designated as the EF-hand, characteristic of the configuration of the metal-ion coordinating ligands of the calcium-binding site. Similarly, the sequence at the amino terminus of the polypeptide of ∼11,000 indicated that it was the light chain counterpart of the same calcium-binding protein complex.",
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T1 - Nuclear proteins of the bovine esophageal epithelium

T2 - I. Monoclonal antibody W2 specifically reacts with condensed nuclei of differentiated superficial cells

AU - Tang, Tang K.

AU - Hong, Tse Ming

AU - Lin, Chen Yong

AU - Lai, Mei Ling

AU - Liu, Catherine H L

AU - Lo, Hsiu Jong

AU - Wang, Ming Eng

AU - Chen, Lan Bo

AU - Chen, Wen Tien

AU - Ip, Wallace

AU - Lin, Diane C.

AU - Lin, Jim J C

AU - Lin, Shin

AU - Sun, Tung Tien

AU - Wang, Eugenia

AU - Wang, John L.

AU - Wu, Reen

AU - Wu, Cheng Wen

AU - Chien, Shu

PY - 1993/2

Y1 - 1993/2

N2 - Cells from three layers of the bovine esophageal epithelium, representing different stages of differentiation, were dissociated and separated by Percoll gradient centrifugation into fractions of small, medium and large sizes. A majority of the large cells possessed condensed nuclei, a characteristic feature of terminal differentiation of the superficial epithelium. The small cells resembled the proliferate cells of the basal layer. In vitro culture of the esophageal epithelial cells resulted in proliferation of the small cells, colony formation, and, in some cases, differentiation into cells with condensed nuclei. Nuclei, or nuclear subfractions derived from cells of the different layers, were used as immunogens for the generation of hybridomas secreting monoclonal antibodies that bound specifically to different regions of the esophageal tissue. One such antibody, designated W2, labeled the condensed nuclei from the superficial layer of stratified esophageal and corneal epithelia in situ, as well as the large cells from esophageal culture in vitro. Thus, the expression of the W2 antigen may be associated with the process of nuclear condensation during epithelial differentiation. Immunoisolation of the target antigen of W2 from extracts of large cells of the bovine esophagus yielded a band of Mr ∼33,000 on nonreducing polyacrylamide gels. This band dissociated into two polypeptides, of Mr ∼22,000 and ∼11,000, upon treatment with dithiothreitol. Amino acid sequence analysis of the larger polypeptide showed extensive homology to a group of small calcium-binding proteins, including two helix-turn-helix motifs designated as the EF-hand, characteristic of the configuration of the metal-ion coordinating ligands of the calcium-binding site. Similarly, the sequence at the amino terminus of the polypeptide of ∼11,000 indicated that it was the light chain counterpart of the same calcium-binding protein complex.

AB - Cells from three layers of the bovine esophageal epithelium, representing different stages of differentiation, were dissociated and separated by Percoll gradient centrifugation into fractions of small, medium and large sizes. A majority of the large cells possessed condensed nuclei, a characteristic feature of terminal differentiation of the superficial epithelium. The small cells resembled the proliferate cells of the basal layer. In vitro culture of the esophageal epithelial cells resulted in proliferation of the small cells, colony formation, and, in some cases, differentiation into cells with condensed nuclei. Nuclei, or nuclear subfractions derived from cells of the different layers, were used as immunogens for the generation of hybridomas secreting monoclonal antibodies that bound specifically to different regions of the esophageal tissue. One such antibody, designated W2, labeled the condensed nuclei from the superficial layer of stratified esophageal and corneal epithelia in situ, as well as the large cells from esophageal culture in vitro. Thus, the expression of the W2 antigen may be associated with the process of nuclear condensation during epithelial differentiation. Immunoisolation of the target antigen of W2 from extracts of large cells of the bovine esophagus yielded a band of Mr ∼33,000 on nonreducing polyacrylamide gels. This band dissociated into two polypeptides, of Mr ∼22,000 and ∼11,000, upon treatment with dithiothreitol. Amino acid sequence analysis of the larger polypeptide showed extensive homology to a group of small calcium-binding proteins, including two helix-turn-helix motifs designated as the EF-hand, characteristic of the configuration of the metal-ion coordinating ligands of the calcium-binding site. Similarly, the sequence at the amino terminus of the polypeptide of ∼11,000 indicated that it was the light chain counterpart of the same calcium-binding protein complex.

KW - Calcium binding protein

KW - Condensed nuclei

KW - Epithelial cell differentiation

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