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The non-glycosylated N-terminal domain of human thrombopoietin is a molten globule under native conditions.
Arai, Shigeki; Shibazaki, Chie; Adachi, Motoyasu; Maeda, Yoshitake; Tahara, Tomoyuki; Kato, Takashi; Miyazaki, Hiroshi; Kuroki, Ryota.
Afiliación
  • Arai S; Quantum Beam Science Research Directorate, National Institutes for Quantum and Radiological Science and Technology (QST), Tokai, Japan.
  • Shibazaki C; Quantum Beam Science Research Directorate, National Institutes for Quantum and Radiological Science and Technology (QST), Tokai, Japan.
  • Adachi M; Quantum Beam Science Research Directorate, National Institutes for Quantum and Radiological Science and Technology (QST), Tokai, Japan.
  • Maeda Y; Kyowa Hakko Kirin Co. Ltd., Tokyo, Japan.
  • Tahara T; Kyowa Hakko Kirin Co. Ltd., Tokyo, Japan.
  • Kato T; Department of Biology, Faculty of Education and Integrated Arts and Sciences, Waseda University, Tokyo, Japan.
  • Miyazaki H; AdipoSeeds, Inc., Tokyo, Japan.
  • Kuroki R; Quantum Beam Science Center, Japan Atomic Energy Agency, Tokai, Japan.
FEBS J ; 286(9): 1717-1733, 2019 05.
Article en En | MEDLINE | ID: mdl-30675759
Human thrombopoietin (hTPO) is a primary hematopoietic growth factor that regulates megakaryocytopoiesis and platelet production. The non-glycosylated form of 1-163 residues of hTPO (hTPO163 ) including the N-terminal active site domain (1-153 residues) is a candidate for treating thrombocytopenia. However, the autoantigenicity level of hTPO163 is higher than that of the full-length glycosylated hTPO (ghTPO332 ). In order to clarify the structural and physicochemical properties of hTPO163 , circular dichroism (CD) and differential scanning calorimetry (DSC) analyses were performed. CD analysis indicated that hTPO163 undergoes an induced-fit conformational change (+19.0% for helix and -16.7% for ß-strand) upon binding to the neutralizing antibody TN1 in a manner similar to the coupled folding and binding mechanism. Moreover, DSC analysis showed that the thermal transition process of hTPO163 is a multistate transition; hTPO163 is thermally stabilized upon receptor (c-Mpl) binding, as indicated with raising the midpoint (Tm ) temperature of the transition by at least +9.5 K. The conformational variability and stability of hTPO163 indicate that hTPO163 exists as a molten globule under native conditions, which may enable the induced-fit conformational change according to the type of ligands (antibodies and receptor). Additionally, CD and computational analyses indicated that the C-terminal domain (154-332 residues) and glycosylation assists the folding of the N-terminal domain. These observations suggest that the antibody affinity and autoantigenicity of hTPO163 might be reduced, if the conformational variability of hTPO163 is restricted by mutation and/or by the addition of C-terminal domain with glycosylation to keep its conformation suitable for the c-Mpl recognition.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Trombopoyetina Límite: Humans Idioma: En Revista: FEBS J Asunto de la revista: BIOQUIMICA Año: 2019 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Trombopoyetina Límite: Humans Idioma: En Revista: FEBS J Asunto de la revista: BIOQUIMICA Año: 2019 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Reino Unido