Abstract
Tumor autocrine motility factor (AMF) has been detected in and purified from serum-free conditioned medium of human HT-1080 fibrosarcoma cells. Under nonreducing conditions, AMF migrates in sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a single band of 55 kDa but under reducing conditions as a band of 64 kDa. Two-dimensional polyacrylamide gel electrophoresis of the purified AMF resolved two groups of polypeptides with isoelectric points of 6.1 and 6.2 (majors), 6.35 and 6.4 (minors). Purified AMF stimulated HT-1080 cell migration in a dose-dependent fashion. The motility stimulation of the fibrosarcoma cells with AMF is associated with the phosphorylation of the AMF receptor, a 78-kDa cell surface glycoprotein (gp78), suggesting protein kinase participation in migratory signal transduction. The gene encoding gp78 was cloned from an HT-1080 fibrosarcoma complementary DNA library. The deduced sequence encodes a polypeptide of 323 amino acids. The nucleotide and predicted amino acid sequence of the gp78 reveals significant homology with the human suppressor/oncogene p53 protein.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Antibodies, Monoclonal
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Base Sequence
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Biomarkers, Tumor / isolation & purification*
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Cell Line
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Cell Movement / drug effects
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Cloning, Molecular
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DNA, Neoplasm / genetics
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DNA, Neoplasm / isolation & purification
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Fibrosarcoma
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Gene Library
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Glucose-6-Phosphate Isomerase
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HeLa Cells / physiology
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Humans
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Molecular Sequence Data
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Neoplasm Proteins / isolation & purification*
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Neoplasm Proteins / metabolism
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Neoplasm Proteins / pharmacology
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RNA, Neoplasm / genetics
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RNA, Neoplasm / isolation & purification
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Receptors, Autocrine Motility Factor
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Receptors, Cell Surface / genetics*
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Receptors, Cytokine*
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Sequence Homology, Nucleic Acid
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Tumor Suppressor Protein p53 / genetics
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Ubiquitin-Protein Ligases
Substances
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Antibodies, Monoclonal
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Biomarkers, Tumor
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DNA, Neoplasm
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Neoplasm Proteins
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RNA, Neoplasm
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Receptors, Cell Surface
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Receptors, Cytokine
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Tumor Suppressor Protein p53
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AMFR protein, human
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Receptors, Autocrine Motility Factor
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Ubiquitin-Protein Ligases
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Glucose-6-Phosphate Isomerase
Associated data
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GENBANK/D10110
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GENBANK/D10111
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GENBANK/M63175
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GENBANK/M63255
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GENBANK/M63926
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GENBANK/M63977
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GENBANK/M63978
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GENBANK/M63997
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GENBANK/M63998
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GENBANK/M64865