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Prepublished online as a Blood First Edition Paper on May 31, 2002; DOI 10.1182/blood-2002-03-0949.
Submitted March 27, 2002
Accepted May 16, 2002
Biochemical characterization of PRV-1, a novel hematopoietic cell surface receptor, which is overexpressed in Polycythemia rubra vera
Steffen Klippel, Elisabeth Strunck, Christian E Busse, Dirk Behringer, and Heike L Pahl*
Department of Exprimental Anaesthesiology, University Hospital Freiburg, Center for Clinical Research, Freiburg, Germany
Department of Hematology/Oncology, University Hospital Freiburg, Freiburg, Germany
* Corresponding author; email: Heike.Pahl{at}klinikum.uni-freiburg.de.
The cDNA for PRV-1, a novel hematopoietic receptor, was recently cloned by virtue of its overexpression in patients with Polycythemia vera. PRV-1 is a member of the uPAR/CD59/Ly6-family of cell surface receptors, which share a common cysteine-rich domain and are tethered to the cell surface via a GPI-link. We have determined the intron/exon structure of the PRV-1 gene and show that the locus is structurally intact in patients with P. vera. Thus, PRV-1 overexpression in these patients is not due to rearrangement or structural alteration of the gene. Northern Blot analysis detects multiple PRV-1 transcripts. Here we show that these transcripts arise from alternative polyadenylation and encode the same protein. Biochemical analysis reveals that PRV-1 is N-glycosylated and embedded in the cell membrane by a lipid anchor, like other members of this family. Moreover, PRV-1 is shed from the cell surface as soluble protein can be detected in cell supernatants. FACS analysis of stably transfected cells revealed that PRV-1 is recognized by antibodies directed against the neutrophil antigen NB1/CD177. Flow cytometry of bone marrow and peripheral blood of both healthy donors and P. vera patients showed that PRV-1 protein is expressed on myeloid cells of the granulocytic lineage. However, unlike the significant difference in PRV-1 expression observed on the mRNA level, the amount of PRV-1 protein on the cell surface is not consistently elevated in P. vera patients compared to healthy controls. Therefore, quantification of PRV-1 surface expression cannot be used for the diagnosis of P. vera.
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