Abstract
Flt4 is a receptor protein tyrosine kinase that is expressed in the adult lymphatic endothelium and high endothelial venules. We have used a BIAcore assay to identify rodent and human cell conditioned media containing the ligand of Flt4 (Flt4-L). Receptor-based affinity chromatography was used to purify this growth factor, followed by amino acid sequencing and molecular cloning of the murine cDNA, the orthologue of human vascular endothelial growth factor-C and vascular endothelial growth factor related protein. The murine flt4-L gene was localized to chromosome 8 and demonstrated to be widely expressed. Flt4-L was found to have a hydrophobic signal sequence and a pro-peptide-like sequence that is removed to generate the mature N-terminus. In addition, the C-terminal region of Flt4-L has four repeats of a cysteine-rich motif that is presumably also proteolytically processed to generate the 21000 Mr polypeptide subunit of the Flt4-L homodimer. Recombinant Flt4-L activated Flt4 as judged by induction of tyrosyl phosphorylation, and induced mitogenesis in vitro of lymphatic endothelial cells.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Animals
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Blotting, Northern
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COS Cells
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Cell Line
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Chromatography, Affinity
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Conserved Sequence
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Culture Media, Conditioned
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Electrophoresis, Polyacrylamide Gel
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Endothelial Growth Factors / metabolism
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Endothelium / drug effects
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Humans
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Immunoglobulin Variable Region / genetics
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Immunoglobulin Variable Region / metabolism
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Mice
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Molecular Sequence Data
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Polymerase Chain Reaction
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Polymorphism, Single-Stranded Conformational
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Rats
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Receptor Protein-Tyrosine Kinases / chemistry
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Receptor Protein-Tyrosine Kinases / genetics*
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Receptor Protein-Tyrosine Kinases / metabolism*
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Receptors, Cell Surface / chemistry
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Receptors, Cell Surface / genetics*
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Receptors, Cell Surface / metabolism*
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Sequence Homology, Amino Acid
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Tissue Distribution
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Transfection
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Vascular Endothelial Growth Factor C
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Vascular Endothelial Growth Factor Receptor-3
Substances
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Culture Media, Conditioned
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Endothelial Growth Factors
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Immunoglobulin Variable Region
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Receptors, Cell Surface
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Recombinant Fusion Proteins
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Vascular Endothelial Growth Factor C
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Receptor Protein-Tyrosine Kinases
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Vascular Endothelial Growth Factor Receptor-3
Associated data
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GENBANK/U58111
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GENBANK/U58112
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GENBANK/X80171