Characterization of a ligand for receptor protein-tyrosine kinase HTK expressed in immature hematopoietic cells.
HTK is a receptor tyrosine kinase that belongs to the Eph subfamily. An extensive screening using BIAcore system revealed that a colon cancer cell line, C-1, expressed the ligand for HTK. From the conditioned medium of C-1 cells, a soluble form of ligand was purified by receptor affinity chromatography, and the isolation of full-length cDNA revealed that this ligand is identical to the human HTK ligand (HTKL) previously reported. HTK receptor tyrosine phosphorylation was induced by membrane-bound or clustered soluble HTKL but not by unclustered soluble HTKL, indicating that HTKL requires cell-to-cell interaction for receptor activation. Binding analysis demonstrated that HTKL binds to HTK with a much higher affinity (Kd: 1.23 nM) than the other transmembrane-type ligand for Eph family, LERK-2/ELKL (Kd: 135 nM). The expression of HTK in cord blood cells was upregulated after the culture in the presence of stem cell factor. Clustered soluble HTKL stimulated the proliferation of sorted HTK+ cord blood cells and a hematopoietic cell line, UT-7/EPO from which HTK was isolated. These findings suggest the involvement of HTK-HTKL system in the proliferation of HTK+ hematopoietic progenitor cells in the hematopoietic environment.
Sakano
S
S
Fundamental Research Laboratory of Life Science, Institute for Life Science Research, Asahi Chemical Industry Co Ltd., Shizuoka-ken, Japan.
Serizawa
R
R
Inada
T
T
Iwama
A
A
Itoh
A
A
Kato
C
C
Shimizu
Y
Y
Shinkai
F
F
Shimizu
R
R
Kondo
S
S
Ohno
M
M
Suda
T
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D036388:Ephrin-B2; D005312:Fetal Blood / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D036224:Receptor, EphB4; D011994:Recombinant Proteins / Q000378:metabolism
D017931:DNA Primers; D018076:DNA, Complementary; D036388:Ephrin-B2; D008024:Ligands; D008565:Membrane Proteins; D011994:Recombinant Proteins; D020794:Receptor Protein-Tyrosine Kinases; D036224:Receptor, EphB4