Purification of bovine S100A12 from recombinant Escherichia coli

Kayoko Yamashita, Yuhta Oyama, Tsuyoshi Shishibori, Osamu Matsushita, Akinobu Okabe, Ryoji Kobayashi

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)47-52
Number of pages6
JournalProtein Expression and Purification
Volume16
Issue number1
DOIs
Publication statusPublished - Jun 1999
Externally publishedYes

Fingerprint

Escherichia coli
Purification
Hypotonic Solutions
EF Hand Motifs
Proteins
Calcium-Binding Proteins
Peptide Mapping
S100 Proteins
Ion Exchange Chromatography
Chromatography
Washing
Edetic Acid
Methionine
Amino Acid Sequence
Ion exchange
Complementary DNA
High Pressure Liquid Chromatography
Cells
Amino Acids
Peptides

Keywords

  • Antiallergy drug
  • Calcium-binding protein
  • Drug affinity chromatography
  • Overexpression
  • Recombinant protein
  • S100 family protein
  • T7 RNA polymerase

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification of bovine S100A12 from recombinant Escherichia coli. / Yamashita, Kayoko; Oyama, Yuhta; Shishibori, Tsuyoshi; Matsushita, Osamu; Okabe, Akinobu; Kobayashi, Ryoji.

In: Protein Expression and Purification, Vol. 16, No. 1, 06.1999, p. 47-52.

Research output: Contribution to journalArticle

Yamashita, K, Oyama, Y, Shishibori, T, Matsushita, O, Okabe, A & Kobayashi, R 1999, 'Purification of bovine S100A12 from recombinant Escherichia coli', Protein Expression and Purification, vol. 16, no. 1, pp. 47-52. https://doi.org/10.1006/prep.1998.1026
Yamashita, Kayoko ; Oyama, Yuhta ; Shishibori, Tsuyoshi ; Matsushita, Osamu ; Okabe, Akinobu ; Kobayashi, Ryoji. / Purification of bovine S100A12 from recombinant Escherichia coli. In: Protein Expression and Purification. 1999 ; Vol. 16, No. 1. pp. 47-52.
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