Purification of human tumor cell autocrine motility factor and molecular cloning of its receptor.
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.
Watanabe
H
H
Cancer Metastasis Program, Michigan Cancer Foundation, Detroit 48201.
Carmi
P
P
Hogan
V
V
Raz
T
T
Silletti
S
S
Nabi
I R
IR
Raz
A
A
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000302:isolation & purification*; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D003001:Cloning, Molecular; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D005354:Fibrosarcoma; D015723:Gene Library; D005956:Glucose-6-Phosphate Isomerase; D006367:HeLa Cells / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D012334:RNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D060285:Receptors, Autocrine Motility Factor; D011956:Receptors, Cell Surface / Q000235:genetics*; D018121:Receptors, Cytokine; D012689:Sequence Homology, Nucleic Acid; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D044767:Ubiquitin-Protein Ligases
D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; D009363:Neoplasm Proteins; D012334:RNA, Neoplasm; D011956:Receptors, Cell Surface; D018121:Receptors, Cytokine; D016159:Tumor Suppressor Protein p53; C555000:AMFR protein, human; D060285:Receptors, Autocrine Motility Factor; D044767:Ubiquitin-Protein Ligases; D005956:Glucose-6-Phosphate Isomerase