Evidence for a precursor of the high-affinity metastasis-associated murine laminin receptor
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Evidence for a precursor of the high-affinity metastasis-associated murine laminin receptor. / Rao, C N; Castronovo, V; Schmitt, M C; Wewer, U M; Claysmith, A P; Liotta, L A; Sobel, M E.
In: Biochemistry, Vol. 28, No. 18, 05.09.1989, p. 7476-86.Research output: Contribution to journal › Journal article › Research › peer-review
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T1 - Evidence for a precursor of the high-affinity metastasis-associated murine laminin receptor
AU - Rao, C N
AU - Castronovo, V
AU - Schmitt, M C
AU - Wewer, U M
AU - Claysmith, A P
AU - Liotta, L A
AU - Sobel, M E
PY - 1989/9/5
Y1 - 1989/9/5
N2 - The high-affinity cellular receptor for the basement membrane component laminin is differentially expressed during tumor invasion and metastasis. A cDNA clone encoding the murine laminin receptor was isolated and identified on the basis of sequence homology to the human laminin receptor [Wewer et al. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 7137-7141]. Primer extension experiments demonstrated that the clone contained the complete 5' sequence of the murine laminin receptor mRNA. RNA blot data demonstrated a single-sized laminin receptor mRNA, approximately 1400 bases long, in human, mouse, and rat. The nascent laminin receptor predicted from the cDNA sequence is 295 amino acids long, with a molecular weight of 33,000, and contains one intradisulfide bridge, a short putative transmembrane domain, and an extracellular carboxy-terminal region which has abundant glutamic acid residues and multiple repeat sequences. The precursor of the laminin receptor is apparently smaller than the 67-kilodalton protein isolated from tissue. The apparent molecular weight on SDS-polyacrylamide gels of the rabbit reticulocyte cell-free translation product of selectively hybridized laminin receptor mRNA is 37,000. Antisera to three different domains of the cDNA-predicted receptor were used to study the relationship between the 37- and 67-kilodalton polypeptides. Antisera to cDNA-deduced synthetic peptides of the receptor immunoprecipitated a 37-kilodalton band both from cell-free translation products and from pulse-labeled cell extracts. On immunoblots of cell extracts, one antisynthetic peptide antiserum recognized only the 67-kilodalton receptor, while another antiserum identified both 37- and 67-kilodalton polypeptides, suggesting a precursor-product relationship between the two polypeptides.
AB - The high-affinity cellular receptor for the basement membrane component laminin is differentially expressed during tumor invasion and metastasis. A cDNA clone encoding the murine laminin receptor was isolated and identified on the basis of sequence homology to the human laminin receptor [Wewer et al. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 7137-7141]. Primer extension experiments demonstrated that the clone contained the complete 5' sequence of the murine laminin receptor mRNA. RNA blot data demonstrated a single-sized laminin receptor mRNA, approximately 1400 bases long, in human, mouse, and rat. The nascent laminin receptor predicted from the cDNA sequence is 295 amino acids long, with a molecular weight of 33,000, and contains one intradisulfide bridge, a short putative transmembrane domain, and an extracellular carboxy-terminal region which has abundant glutamic acid residues and multiple repeat sequences. The precursor of the laminin receptor is apparently smaller than the 67-kilodalton protein isolated from tissue. The apparent molecular weight on SDS-polyacrylamide gels of the rabbit reticulocyte cell-free translation product of selectively hybridized laminin receptor mRNA is 37,000. Antisera to three different domains of the cDNA-predicted receptor were used to study the relationship between the 37- and 67-kilodalton polypeptides. Antisera to cDNA-deduced synthetic peptides of the receptor immunoprecipitated a 37-kilodalton band both from cell-free translation products and from pulse-labeled cell extracts. On immunoblots of cell extracts, one antisynthetic peptide antiserum recognized only the 67-kilodalton receptor, while another antiserum identified both 37- and 67-kilodalton polypeptides, suggesting a precursor-product relationship between the two polypeptides.
KW - Amino Acid Sequence
KW - Animals
KW - Base Sequence
KW - DNA
KW - Humans
KW - Immunoblotting
KW - Laminin
KW - Mice
KW - Molecular Sequence Data
KW - Neoplasm Metastasis
KW - Nucleic Acid Conformation
KW - Nucleic Acid Hybridization
KW - Protein Biosynthesis
KW - Protein Conformation
KW - Protein Precursors
KW - RNA, Messenger
KW - Rats
KW - Receptors, Immunologic
KW - Receptors, Laminin
KW - Restriction Mapping
KW - Sequence Homology, Nucleic Acid
KW - Tumor Cells, Cultured
M3 - Journal article
C2 - 2531008
VL - 28
SP - 7476
EP - 7486
JO - Biochemistry
JF - Biochemistry
SN - 0006-2960
IS - 18
ER -
ID: 34330128