Ein allgemeines Selektionssystem zur Stabilisierung von Proteinen

• Antragsteller: Professor Dr. Franz-Xaver Schmid
• Fachliche Zuordnung: Biochemie
• Förderung: Förderung von 2000 bis 2008
• Projektkennung: Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 5301648

• Erstellungsjahr: 2011

Zusammenfassung der Projektergebnisse

Durch in-vitro Selektion konnten für mehrere Proteine eine Vielzahl stark stabilisierter Varianten zu erzeugen. Die eingehende Analyse der Ergebnisse in Bezug auf die Stabilität und die Struktur der optimierten Proteinvarianten hat neue Einsichten in die molekularen Grundlagen der Proteinstabilität geliefert.

Projektbezogene Publikationen (Auswahl)

• (2001). In-vitro selection of highly stabilized protein variants with optimized surface. J. Mol. Biol. 309, 717-726
  Martin, A., Sieber, V., and Schmid, F. X.
  
• (2002). Origins of the high stability of an in vitro-selected cold-shock protein. J. Mol. Biol. 318, 1341-1349
  Martin, A., Kather, I., and Schmid, F. X.
  
• (2003). A proline switch controls folding and domain interactions in the gene-3-protein of the filamentous phage fd. J. Mol. Biol. 331, 1131-1140
  Martin, A., and Schmid, F. X.
  
• (2003). Evolutionary stabilization of the gene-3-protein of phage fd reveals the principles that govern the thermodynamic stability of two-domain proteins. J. Mol. Biol. 328, 863-875
  Martin, A., and Schmid, F. X.
  
• (2003). Proside: a phage based method for selecting thermostable proteins. Methods in Molecular Biology 230, 57-70
  Martin, A., Schmid, F. X., and Sieber, V.
  
• (2003). The folding mechanism of a two-domain protein: folding kinetics and domain docking of the gene-3-protein of phage fd. J. Mol. Biol. 329, 599-610
  Martin, A., and Schmid, F. X.
  
• (2005). A stable disulfide-free gene-3-protein of phage fd generated by in vitro evolution. J. Mol. Biol. 354, 666-678
  Kather, I., Bippes, C. A., and Schmid, F. X.
  
• (2005). Evolutionary protein stabilization in comparison with computational design. J. Mol. Biol. 351, 1160-1168
  Wunderlich, M., Martin, A., Staab, C. A., and Schmid, F. X.
  
• (2005). Stabilization of the cold shock protein CspB from Bacillus subtilis by evolutionary optimization of Coulombic interactions. J. Mol. Biol. 347, 1063-1076
  Wunderlich, M., Martin, A., and Schmid, F. X.
  
• (2006). In vitro evolution of a hyperstable Gbeta1 variant. J. Mol. Biol. 363, 545-557
  Wunderlich, M., and Schmid, F. X.
  
• (2006). The correlation between protein stability and dipole moment: a critical test. Protein. Eng. Des. Sel. 19, 355-358
  Wunderlich, M., and Schmid, F. X.
  
• (2007). Optimization of the G-beta1 domain by computational design and by in vitro evolution: structural and energetic basis of stabilization. J. Mol. Biol. 373, 775-784
  Wunderlich, M., Max, K. E., Roske, Y., Mueller, U., Heinemann, U., and Schmid, F. X.
  
• (2007). Optimized variants of the cold shock protein from in vitro selection: structural basis of their high thermostability. J. Mol. Biol. 369, 1087-1097
  Max, K. E., Wunderlich, M., Roske, Y., Schmid, F. X., and Heinemann, U.
  
• (2008). Changing the determinants of protein stability from covalent to non-covalent interactions by in-vitro evolution: a structural and energetic analysis. J. Mol. Biol. 381, 1040-1054
  Kather, I., Jakob, R., Dobbek, H., and Schmid, F. X.
  
• (2008). Increased Folding Stability of TEM-1 beta-Lactamase by In Vitro Selection. J. Mol. Biol. 383, 238-251
  Kather, I., Jakob, R. P., Dobbek, H., and Schmid, F. X.