Functional characterization of c-Mpl ectodomain mutations that underlie congenital amegakaryocytic thrombocytopenia
Details
Publication Year 2014-01-20,Volume 32,Issue #1,Page 18-26
Journal Title
Growth Factors
Publication Type
Journal Article
Abstract
Abstract Activation of the cell surface receptor, c-Mpl, by the cytokine, thrombopoietin (TPO), underpins megakaryocyte and platelet production in mammals. In humans, mutations in c-Mpl have been identified as the molecular basis of Congenital Amegakaryocytic Thrombocytopenia (CAMT). Here, we show that CAMT-associated mutations in c-Mpl principally lead to defective receptor presentation on the cell surface. In contrast, one CAMT mutant c-Mpl, F104S, was expressed on the cell surface, but showed defective TPO binding and receptor activation. Using mutational analyses, we examined which residues adjacent to F104 within the membrane-distal cytokine receptor homology module (CRM) of c-Mpl comprise the TPO-binding epitope, revealing residues within the predicted Domain 1 E-F and A-B loops and Domain 2 F'-G' loop as key TPO-binding determinants. These studies underscore the importance of the c-Mpl membrane-distal CRM to TPO-binding and suggest that mutations within this CRM that perturb TPO binding could give rise to CAMT.
Publisher
Informa
Keywords
Congenital amegakaryocytic thrombocytopenia, ligand binding, Mpl, TPO47
Research Division(s)
Cancer And Haematology; Molecular Medicine; Structural Biology
NHMRC Grants
NHMRC/1016647
Terms of Use/Rights Notice
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Creation Date: 2014-01-24 12:02:24
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