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| SHP-2 protein, which has two SH2 domains, is essential for the embryonic development, haematopoiesis and signaling downstream of a variety of growth factors. SHP-2 proteins are related to many diseases. To facilitate fundamental studies, it is important that the proteins can be expressed in high quality and in a large quantity. In this work, the amino-terminal SH2 (NSH2) domain of SHP-2 protein which is important for the protein’s self-regulation was recombinated into the prokaryotic expression vector, pGEX-2T, and introduced into E. coli BL21 for a prokaryotic expression. Two recombinant vectors with different extra amino acid residues were designed on the basis of properties of NSH2 protein and the sequence of expression vector, pGEX-2T, to produce NSH2 protein with a high property. The effects of the two groups of different extra amino acid residues on solubility and stability were compared. The comparisons indicated that end extra amino acid residues may have strong effects on both solubility and stability. The higher soluble and stable NSH2 protein was selected and labeled with isotope 15N for NMR study. The high resolution of NMR demonstrated the correction folding of the protein and the interactions of the protein with phosphop-tyrosin peptides. |
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| Keywords:SHP-2; SH2; protein purification |
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