Identification of fetomodulin, a surface marker protein of fetal development, as thrombomodulin by gene cloning and functional assays.
Fetomodulin (FM) was previously shown to be a surface marker protein of parietal endoderm by in vitro differentiation of F9 embryonal carcinoma cells and by immunohistochemistry of in vivo embryos. BALB/3T3 and sarcoma S180 cells of the mouse were also shown to possess a protein which was indistinguishable from FM by immunological and structural criteria. We now show by protein and DNA sequencing and by functional assays that FM is identical to thrombomodulin, an anticoagulant endothelial thrombin receptor. Partial amino acid sequences of FM from S180 cells suggested homology between FM and thrombomodulin. An FM cDNA fragment was obtained by screening an expression library, which was constructed with restricted BALB/3T3 cDNA, with polyclonal anti-FM antibody. Several longer cDNA clones were than isolated using this fragment as a probe. They elucidated a 3369-bp partial sequence which encompassed 93% of the coding sequence. The remaining structure was determined from a genomic DNA clone. The deduced FM structure proved to be identical to that of thrombomodulin of mouse lung. Affinity-purified FM of BALB/3T3 and differentiated F9 cells was as active as thrombomodulin of the lung in binding thrombin and also as an anticoagulant. Structural and functional identity of the two proteins was thus confirmed. During embryonic development, FM immunoreactivity is localized not only in vasculatures but also at sites of cell-to-cell contact, including lung bud and neural epithelium, which were not expected a priori to possess this endothelial surface protein. FM may be a multifunctional protein with unique roles in embryonic development.
Imada
S
S
Division of Cell Biology, Meiji Institute of Health Science, Meiji Milk Products Co. Ltd., Odawara, Japan.
Yamaguchi
H
H
Nagumo
M
M
Katayanagi
S
S
Iwasaki
H
H
Imada
M
M
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D002454:Cell Differentiation; D003001:Cloning, Molecular; D004848:Epithelium / Q000378:metabolism; D066298:In Vitro Techniques; D008168:Lung / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000032:analysis / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D011486:Protein C / Q000096:biosynthesis; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000502:physiology; D018179:Receptors, Thrombin; D013917:Thrombin / Q000378:metabolism
C052976:FM protein, mouse; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011486:Protein C; D011956:Receptors, Cell Surface; D018179:Receptors, Thrombin; D013917:Thrombin