Chakravarty, Suvobrata and Mitra, Nivedita and Queitsch, Iris and Surolia, Avadhesha and Varadarajan, Raghavan and Dubel, Stefan (2000) Protein stabilization through phage display. In: FEBS Letters, 476 (3). pp. 296-300.
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RNase S consists of two proteolytic fragments of RNase A, residues 1–20 (S20) and residues 21–124 (S pro). A 15-mer peptide (S15p) with high affinity for S pro was selected from a phage display library. Peptide residues that are buried in the structure of the wild type complex are conserved in S15p though there are several changes at other positions. Isothermal titration calorimetry studies show that the affinity of S15p is comparable to that of the wild type peptide at $25^0C$. However, the magnitudes of $\Delta H^0$ and $\Delta C_p$ are lower for S15p, suggesting that the thermal stability of the complex is enhanced. In agreement with this prediction, at pH 6, the $T_m$ of the S15p complex was found to be $10^0C$ higher than that of the wild type complex. This suggests that for proteins where fragment complementation systems exist, phage display can be used to find mutations that increase protein thermal stability.
|Item Type:||Journal Article|
|Additional Information:||Copyright of this article belongs to Federation of European Biochemical Societies|
|Keywords:||Phage display;Fragment complementation;Thermal stability;Epitope mapping|
|Department/Centre:||Division of Biological Sciences > Molecular Biophysics Unit|
|Date Deposited:||04 Oct 2007|
|Last Modified:||19 Sep 2010 04:40|
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