Cleavage of recombinant proteins at poly-His sequences by Co(II)and Cu(II)

M. Andberg, J. Jäntti, S. Heilimo, P. Pihkala, A. Paananen, A.M.P. Koskinen, H. Söderlund, M.B. Linder

Research output: Contribution to journalArticleScientificpeer-review

14 Citations (Scopus)

Abstract

Improved ways to cleave peptide chains at engineered sites easily and specifically would form useful tools for biochemical research. Uses of such methods include the activation or inactivation of enzymes or the removal of tags for enhancement of recombinant protein expression or tags used for purification of recombinant proteins. In this work we show by gel electrophoresis and mass spectroscopy that salts of Co(II) and Cu(II) can be used to cleave fusion proteins specifically at sites where sequences of His residues have been introduced by protein engineering. The His residues could be either consecutive or spaced with other amino acids in between. The cleavage reaction required the presence of low concentrations of ascorbate and in the case of Cu(II) also hydrogen peroxide. The amount of metal ions required for cleavage was very low; in the case of Cu(II) only one to two molar equivalents of Cu(II) to protein was required. In the case of Co(II), 10 molar equivalents gave optimal cleavage. The reaction occurred within minutes, at a wide pH range, and efficiently at temperatures ranging from 0°C to 70°C. The work described here can also have implications for understanding protein stability in vitro and in vivo.
Original languageEnglish
Pages (from-to)1751-1761
JournalProtein Science
Volume16
Issue number8
DOIs
Publication statusPublished - 2007
MoE publication typeA1 Journal article-refereed

Fingerprint

Recombinant Proteins
Protein Engineering
Protein Stability
Hydrogen Peroxide
Electrophoresis
Mass Spectrometry
Proteins
Salts
Gels
Metals
Ions
Amino Acids
Peptides
Temperature
Enzymes
Research
Purification
Metal ions
Fusion reactions
Chemical activation

Keywords

  • protein cleavage
  • His tag
  • artificial protease
  • protein stability
  • Co(II)
  • Cu(II)

Cite this

Andberg, M. ; Jäntti, J. ; Heilimo, S. ; Pihkala, P. ; Paananen, A. ; Koskinen, A.M.P. ; Söderlund, H. ; Linder, M.B. / Cleavage of recombinant proteins at poly-His sequences by Co(II)and Cu(II). In: Protein Science. 2007 ; Vol. 16, No. 8. pp. 1751-1761.
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Cleavage of recombinant proteins at poly-His sequences by Co(II)and Cu(II). / Andberg, M.; Jäntti, J.; Heilimo, S.; Pihkala, P.; Paananen, A.; Koskinen, A.M.P.; Söderlund, H.; Linder, M.B.

In: Protein Science, Vol. 16, No. 8, 2007, p. 1751-1761.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Jäntti, J.

AU - Heilimo, S.

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AU - Söderlund, H.

AU - Linder, M.B.

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