TY - JOUR
T1 - Purification of bovine S100A12 from recombinant Escherichia coli
AU - Yamashita, Kayoko
AU - Oyama, Yuhta
AU - Shishibori, Tsuyoshi
AU - Matsushita, Osamu
AU - Okabe, Akinobu
AU - Kobayashi, Ryoji
N1 - Funding Information:
We thank Yasuharu Sasaki at the Asahi-kasei Life Science General Institute, Shizuoka, Japan, for technical assistance with peptide sequencing. This work was supported in part by Grants-in-Aid from the Fugaku Trust for Medical Research (to R.K.) and for Scientific Research (B) from The Ministry of Education, Science, Sports and Culture of Japan (to R.K.).
PY - 1999/6
Y1 - 1999/6
N2 - S100A12, a member of the S100 family of EF-hand calcium-binding proteins, was purified from Escherichia coli cells expressing the corresponding cDNA. The procedure involved washing induced E. coli cells with EDTA-containing hypotonic solution, ion-exchange chromatography, and HPLC. Recombinant S100A12 was purified to homogeneity with the final yield around 6.7 mg per 20 ml of culture. The purified protein was identical to native S100A12 in the N-terminal amino acid sequence, lysylendopeptidase peptide mapping, mass spectrum, and Ca2+-dependent binding affinity to amlexanox, an antiallergy drug. However, the N-terminal methionine residue of the purified protein was not cleaved off as in the native protein. The method used in the present study permits the purification of recombinant S100A12 in large quantities and may also be applicable to preparation of other S100 family proteins.
AB - S100A12, a member of the S100 family of EF-hand calcium-binding proteins, was purified from Escherichia coli cells expressing the corresponding cDNA. The procedure involved washing induced E. coli cells with EDTA-containing hypotonic solution, ion-exchange chromatography, and HPLC. Recombinant S100A12 was purified to homogeneity with the final yield around 6.7 mg per 20 ml of culture. The purified protein was identical to native S100A12 in the N-terminal amino acid sequence, lysylendopeptidase peptide mapping, mass spectrum, and Ca2+-dependent binding affinity to amlexanox, an antiallergy drug. However, the N-terminal methionine residue of the purified protein was not cleaved off as in the native protein. The method used in the present study permits the purification of recombinant S100A12 in large quantities and may also be applicable to preparation of other S100 family proteins.
KW - Antiallergy drug
KW - Calcium-binding protein
KW - Drug affinity chromatography
KW - Overexpression
KW - Recombinant protein
KW - S100 family protein
KW - T7 RNA polymerase
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U2 - 10.1006/prep.1998.1026
DO - 10.1006/prep.1998.1026
M3 - Article
C2 - 10336859
AN - SCOPUS:0033153513
SN - 1046-5928
VL - 16
SP - 47
EP - 52
JO - Protein Expression and Purification
JF - Protein Expression and Purification
IS - 1
ER -