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 language | English (US) |
|---|---|
| Pages (from-to) | 571-578 |
| Number of pages | 8 |
| Journal | Journal of Cellular Physiology |
| Volume | 164 |
| Issue number | 3 |
| DOIs | |
| State | Published - Sep 1995 |
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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 journal › Article
}
TY - JOUR
T1 - Two nuclear proteins in tracheal epithelial cells are recognized by antibodies specific to a squamous differentiation marker, sPRI
AU - Tesfaigzi, Johannes
AU - An, Gang
AU - Wu, Reen
AU - Carlson, Don M.
PY - 1995/9
Y1 - 1995/9
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0029086822&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0029086822&partnerID=8YFLogxK
U2 - 10.1002/jcp.1041640315
DO - 10.1002/jcp.1041640315
M3 - Article
C2 - 7650064
AN - SCOPUS:0029086822
VL - 164
SP - 571
EP - 578
JO - Journal of Cellular Physiology
JF - Journal of Cellular Physiology
SN - 0021-9541
IS - 3
ER -