共 35 条
Protein stabilization through phage display
被引:7
作者:
Chakravarty, S
Mitra, N
Queitsch, I
Surolia, A
Varadarajan, R
[1
]
Dübel, S
机构:
[1] Indian Inst Sci, Mol Biophys Unit, Bangalore 560012, Karnataka, India
[2] Heidelberg Univ, D-69120 Heidelberg, Germany
[3] Indian Inst Sci, Jawaharlal Nehru Ctr Adv Sci Res, Chem Biol Unit, Bangalore 560012, Karnataka, India
关键词:
phage display;
fragment complementation;
thermal stability;
epitope mapping;
D O I:
10.1016/S0014-5793(00)01725-7
中图分类号:
Q5 [生物化学];
Q7 [分子生物学];
学科分类号:
071010 ;
081704 ;
摘要:
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 degrees C. However, the magnitudes of Delta H degrees and Delta C-p are lower for S15p, suggesting that the thermal stability of the complex is enhanced. In agreement with this prediction, at pit 6, the T-m of the S15p complex was found to he 10 degrees C 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. (C) 2000 Federation of European Biochemical Societies, Published by Elsevier Science B.V. All rights reserved.
引用
收藏
页码:296 / 300
页数:5
相关论文