Characterization of the protein encoded by the flt3 (flk2) receptor-like tyrosine kinase gene.
We have developed rabbit polyclonal antibodies to the C-terminus of the flt3-encoded protein, which is a member of the receptor tyrosine kinase family. Immunoprecipitation using this antiserum brings down two protein bands, a major band of 143 kDa and a less abundant, more diffuse, band of 158 kDa. Pulse-chase analysis of flt3 protein from transfected COS-7 cells shows that the larger band is derived from the smaller one and presumably represents maturation of the protein from a glycosylated high-mannose form to a complex carbohydrate form. N-glycosidase F digestion confirmed the presence of N-linked carbohydrates, and cell-surface labeling of flt3-transfected cells indicated that the 158-kDa glycoprotein is the species found on the cell surface. A mutated form of the flt3 protein that was defective in its glycosylational processing was identified. Western blotting of the immunoprecipitated flt3 protein showed that it is heavily phosphorylated on tyrosine, and that this phosphorylation probably occurs in the absence of ligand. In this regard, the flt3 protein resembles the c-erbB2 protein, which is also highly phosphorylated in the absence of ligand. These data suggest that the flt3 receptor regulates the growth and differentiation of cells via an as yet unknown ligand.
Lyman
S D
SD
Department of Molecular Biology, Immunex Research and Development Corporation, Seattle, Washington 98101.
James
L
L
Zappone
J
J
Sleath
P R
PR
Beckmann
M P
MP
Bird
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D001483:Base Sequence; D002462:Cell Membrane / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D010455:Peptides / Q000276:immunology; D019000:Phosphotyrosine; D011505:Protein-Tyrosine Kinases / Q000737:chemistry* / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000737:chemistry* / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D014443:Tyrosine / Q000031:analogs & derivatives / Q000737:chemistry; D051941:fms-Like Tyrosine Kinase 3
D000906:Antibodies; D006023:Glycoproteins; D009838:Oligodeoxyribonucleotides; D010455:Peptides; D011518:Proto-Oncogene Proteins; D011956:Receptors, Cell Surface; D019000:Phosphotyrosine; D014443:Tyrosine; C497971:Flt3 protein, mouse; D011505:Protein-Tyrosine Kinases; D020794:Receptor Protein-Tyrosine Kinases; D051941:fms-Like Tyrosine Kinase 3; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase