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Multiple conformational states of a new hematopoietic cytokine (megakaryocyte growth and development factor): pH- and urea-induced denaturation.
Hamburger, J B; Chen, E; Narhi, L O; Wu, G M; Brems, D N.
Afiliación
  • Hamburger JB; Amgen Inc., Thousand Oaks, California 91320, USA.
Proteins ; 32(4): 495-503, 1998 Sep 01.
Article en En | MEDLINE | ID: mdl-9726418
The effect of pH and urea on the conformation of recombinant human megakaryocyte growth and development factor (rHuMGDF) was determined by circular dichroism, intrinsic fluorescence spectroscopy, and equilibrium ultracentrifugation. The conformation of rHuMGDF was dependent on pH and urea concentration. Multiple folding forms were evidenced by multiple pH-induced transitions and urea-induced equilibrium transitions that deviated from a simple two-state process. In neutral to alkaline pH, rHuMGDF exists as a monomer, but an acid-induced conformational state self-associates to form a soluble aggregate. A folding intermediate(s) was observed with a more stable secondary structure than tertiary structure and was dependent on the pH of the urea-induced denaturation. The differences in the stabilities of the folding states were most distinct in the pH range of 4.5 to 6.5. The presence of intermediates in the folding pathway of rHuMGDF are similar to findings of previous studies of related growth factors that share a common three-dimensional structure.
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Trombopoyetina Límite: Humans Idioma: En Revista: Proteins Asunto de la revista: BIOQUIMICA Año: 1998 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Trombopoyetina Límite: Humans Idioma: En Revista: Proteins Asunto de la revista: BIOQUIMICA Año: 1998 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos