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Serum form of the erythropoietin receptor identified by a sequence-
specific peptide antibody [see comments]
RD Baynes, GK Reddy, YJ Shih, BS Skikne and JD Cook
Department of Medicine, Kansas University Medical Center, Kansas City
66160-7402.
The present investigation was undertaken to search for soluble forms of the
erythropoietin receptor in human serum using polyclonal antibody against an
amino terminal peptide sequence in the extracellular domain. This sequence
was located adjacent to the amino terminus at residues 25- 38. When this
antibody was used for Western blots of solubilized membranes from nucleated
bone marrow cells, a protein consistent with native erythropoietin receptor
was seen. Purified soluble ectodomain of the erythropoietin receptor
displayed appropriate reactivity with this antibody. When sera from normal
subjects and patients with a range of hematologic disorders were examined
by Western blotting, a protein with a molecular mass of 34 Kd was detected
in sera from patients with enhanced erythropoiesis including sickle cell
anemia, thalassemia, and megaloblastic anemia. This protein was rarely
detected in normal serum but appeared when normal subjects were treated
with recombinant erythropoietin and disappeared after full treatment of
patients with megaloblastic anemia due to vitamin B12 deficiency. The
protein was not detected after myeloablation for bone marrow
transplantation but appeared with marrow engraftment. Reactivity of this
protein with the peptide antibody was competitively inhibited by the amino
terminal peptide sequence. An additional 48 Kd protein was detected that
showed minimal variation in intensity with differing degrees of
erythropoietic activity. Detection of this protein could not be inhibited
by the addition of synthetic peptide. Our findings indicate the presence of
a soluble form of the erythropoietin receptor related to the extracellular
domain that is highly correlated with enhanced erythropoiesis.
Volume 82,
Issue 7,
pp. 2088-2095,
10/01/1993
Copyright © 1993 by The American Society of Hematology
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