Expression and purification of 15N-labeled 2-SH2 protein domain of SHP-2 from Homo sapiens in Escherichia coli for NMR studies and applications.
A method for 2-SH2 protein domain study was described as per the order of expression, purification and structural detection. The 2-SH2 protein of Homo sapiens SHP-2 was successfully expressed and purified. It could specifically bind to anti-SHP-2/SHPTP-2 antibody according to the MS and Western blot analysis. The NMR spectrum result reveals that the protein exists in a well-ordered structure. This can provide foundations to find out the reaction mechanism of the D phosphorylated-EPIYA motif accessible to 2-SH2, support the research and development of the novel detection chip as well as target inhibition medicine for the future clinical applications.
Wu
Ye
Y
Institute of Bioengineering, Zhejiang Sci-Tech University, Xiasha Road #2, Hangzhou, PR China. wuye83.lj@163.com
Guo
Jiang-Feng
JF
D000595:Amino Acid Sequence; D000906:Antibodies / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D017434:Protein Structure, Tertiary / Q000235:genetics*; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11 / Q000302:isolation & purification* / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D014961:X-Ray Diffraction
D000906:Antibodies; D017931:DNA Primers; C516724:PTPN11 protein, human; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11