Two nuclear proteins in tracheal epithelial cells are recognized by antibodies specific to a squamous differentiation marker, sPRI

Johannes Tesfaigzi, Gang An, Reen Wu, Don M. Carlson

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Abstract

In cell-free translations of RNA from primary cultures of pig trachea surface epithelial cells we observed that a mRNA encoding a 20 kDa pro[ine-rich protein (sPRP) was dramatically induced during culturing (Tesfaigzi et al., 1990, Biochem. Biophys. Res. Commun., 172:M1304-1309). This mRNA was not detected in tracheal tissue or in epithelial cells prior to culturing. Antisera were raised to synthetic peptide sequences corresponding to 23 amino acids on the C-terminus (C23-antiserum) and 29 amino acids on the N-terminus (N29 antiserum) of sPRP. On Western blot analysis, C23 antiserum reacted with a 20 kDa protein in cytosolic extracts from pig tracheal cells maintained in culture for 4 days. The reaction with the 20 kDa protein was inhibited by adding C23 peptide. Two nuclear proteins (66 and 70 kDa) obtained by micrococcal nuclease treatment of tracheal cell nuclei were detected on Western blots with C23 antiserum. These proteins were present in cells both before and after culturing. Sucrose gradient fractionation indicated that these nuclear proteins are associated with chromatin. Small amounts of the 66 and 70 kDa proteins were obtained from nuclear matrix fractions. These nuclear proteins also reacted with N29 antiserum. Since these proteins share similar epitopes with the N- and C-termini of sPRP, it is likely that the 20 kDa protein (sPRP) is part of these proteins. However, purification of the nuclear proteins followed by an amino acid sequence analysis is necessary to clarify whether sPRP is part of these proteins.

Original languageEnglish (US)
Pages (from-to)571-578
Number of pages8
JournalJournal of Cellular Physiology
Volume164
Issue number3
DOIs
StatePublished - Sep 1995

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Differentiation Antigens
Nuclear Proteins
Epithelial Cells
Antibodies
Immune Sera
Proteins
Amino Acids
Swine
Western Blotting
Micrococcal Nuclease
Nuclear Matrix
Messenger RNA
Peptides
Protein Sequence Analysis
Fractionation
Trachea
Cell Nucleus
Chromatin
Purification
Sucrose

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Two nuclear proteins in tracheal epithelial cells are recognized by antibodies specific to a squamous differentiation marker, sPRI. / Tesfaigzi, Johannes; An, Gang; Wu, Reen; Carlson, Don M.

In: Journal of Cellular Physiology, Vol. 164, No. 3, 09.1995, p. 571-578.

Research output: Contribution to journalArticle

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