Thrombomodulin is preferentially expressed in Balb/c lung microvessels.
Previously, two rat monoclonal antibodies where developed which bind distinct epitopes on a murine glycoprotein, P112, which is expressed primarily in lung capillary endothelium. In this paper we show that P112 is identical to the endothelial anticoagulant protein, thrombomodulin (TM). Several lines of evidence support this conclusion. First, amino acid analysis of P112 shows a high degree of homology to TM, and both molecules exhibit the same mobility in gel electrophoresis. Second, P112 and TM share reactivity for two different monoclonal antibodies. Third, purified P112, like TM, acts as a cofactor for protein C activation. Finally, two cDNA clones identified with P112 polyclonal antiserum contain sequence identity with the known TM cDNA sequence. Quantitative analysis of TM (P112) expression using a two-site monoclonal antibody assay demonstrates that significantly higher levels of TM are found in lung in comparison with other highly vascularized organs, i.e. the kidney and liver. Quantitative Northern blot data coincides with the two-site assay data and demonstrates that the high level of TM expression in lung is not due to preferential binding of the monoclonal antibodies to lung TM but rather to increased production of TM mRNA in the lung relative to other highly vascularized organs. It is suggested that expression of TM is highest in cells from continuous endothelium.
Oak Ridge Graduate School of Biomedical Sciences, University of Tennessee, Oak Ridge.
V A
VA
Ford
C
C
Stringer
S J
SJ
Kennel
D000818:Animals; D000911:Antibodies, Monoclonal; D015152:Blotting, Northern; D002196:Capillaries / Q000502:physiology*; D004247:DNA / Q000235:genetics; D004730:Endothelium, Vascular / Q000502:physiology*; D005260:Female; D007668:Kidney / Q000502:physiology; D008168:Lung / Q000098:blood supply*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008833:Microcirculation / Q000502:physiology*; D009928:Organ Specificity; D011061:Poly A / Q000235:genetics / Q000302:isolation & purification; D011486:Protein C / Q000378:metabolism; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D051381:Rats; D011956:Receptors, Cell Surface / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D018179:Receptors, Thrombin
D000911:Antibodies, Monoclonal; D011486:Protein C; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D018179:Receptors, Thrombin; D011061:Poly A; D012313:RNA; D004247:DNA