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

T. K. Tang, Tse-Ming Hong, C. Y. Lin, M. L. Lai, C. H.L. Liu, H. J. Lo, M. E. Wang, L. B. Chen, W. T. Chen, W. Ip, D. C. Lin, J. J.C. Lin, S. Lin, T. T. Sun, E. Wang, J. L. Wang, R. Wu, C. W. Wu, S. Chien

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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 laver 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 M(r) ~33,000 on nonreducing polyacrylamide gels. This band dissociated into two polypeptides, of M(r) ~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
Pages (from-to)237-247
Number of pages11
JournalJournal of Cell Science
Volume104
Issue number2
Publication statusPublished - 1993 Jan 1

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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

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

Tang, T. K. ; Hong, Tse-Ming ; Lin, C. Y. ; Lai, M. L. ; Liu, C. H.L. ; Lo, H. J. ; Wang, M. E. ; Chen, L. B. ; Chen, W. T. ; Ip, W. ; Lin, D. C. ; Lin, J. J.C. ; Lin, S. ; Sun, T. T. ; Wang, E. ; Wang, J. L. ; Wu, R. ; Wu, C. W. ; Chien, S. / 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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title = "Nuclear proteins of the bovine esophageal epithelium. I. Monoclonal antibody W2 specifically reacts with condensed nuclei of differentiated superficial cells",
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 laver 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 M(r) ~33,000 on nonreducing polyacrylamide gels. This band dissociated into two polypeptides, of M(r) ~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.",
author = "Tang, {T. K.} and Tse-Ming Hong and Lin, {C. Y.} and Lai, {M. L.} and Liu, {C. H.L.} and Lo, {H. J.} and Wang, {M. E.} and Chen, {L. B.} and Chen, {W. T.} and W. Ip and Lin, {D. C.} and Lin, {J. J.C.} and S. Lin and Sun, {T. T.} and E. Wang and Wang, {J. L.} and R. Wu and Wu, {C. W.} and S. Chien",
year = "1993",
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Tang, TK, Hong, T-M, 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.

Nuclear proteins of the bovine esophageal epithelium. I. Monoclonal antibody W2 specifically reacts with condensed nuclei of differentiated superficial cells. / Tang, T. K.; Hong, Tse-Ming; Lin, C. Y.; Lai, M. L.; Liu, C. H.L.; Lo, H. J.; Wang, M. E.; Chen, L. B.; Chen, W. T.; Ip, W.; Lin, D. C.; Lin, J. J.C.; Lin, S.; Sun, T. T.; Wang, E.; Wang, J. L.; Wu, R.; Wu, C. W.; Chien, S.

In: Journal of Cell Science, Vol. 104, No. 2, 01.01.1993, p. 237-247.

Research output: Contribution to journal › Article

TY - JOUR

T1 - Nuclear proteins of the bovine esophageal epithelium. I. Monoclonal antibody W2 specifically reacts with condensed nuclei of differentiated superficial cells

AU - Tang, T. K.

AU - Hong, Tse-Ming

AU - Lin, C. Y.

AU - Lai, M. L.

AU - Liu, C. H.L.

AU - Lo, H. J.

AU - Wang, M. E.

AU - Chen, L. B.

AU - Chen, W. T.

AU - Ip, W.

AU - Lin, D. C.

AU - Lin, J. J.C.

AU - Lin, S.

AU - Sun, T. T.

AU - Wang, E.

AU - Wang, J. L.

AU - Wu, R.

AU - Wu, C. W.

AU - Chien, S.

PY - 1993/1/1

Y1 - 1993/1/1

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 laver 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 M(r) ~33,000 on nonreducing polyacrylamide gels. This band dissociated into two polypeptides, of M(r) ~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 laver 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 M(r) ~33,000 on nonreducing polyacrylamide gels. This band dissociated into two polypeptides, of M(r) ~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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M3 - Article

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VL - 104

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JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

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Tang TK, Hong T-M, Lin CY, Lai ML, Liu CHL, Lo HJ et al. Nuclear proteins of the bovine esophageal epithelium. I. Monoclonal antibody W2 specifically reacts with condensed nuclei of differentiated superficial cells. Journal of Cell Science. 1993 Jan 1;104(2):237-247.

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