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

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dc.contributor Aalto-yliopisto fi
dc.contributor Aalto University en Anberg, Martina Jäntti, Jussi Heilimo, Saara Pihkala, Päivi Paananen, Arja Koskinen, Ari M.P. Söderlund, Hans Linder, Markus 2016-09-23T07:39:25Z 2007-08
dc.identifier.citation Anberg , M , Jäntti , J , Heilimo , S , Pihkala , P , Paananen , A , Koskinen , A M P , Söderlund , H & Linder , M 2007 , ' Cleavage of recombinant proteins at poly-His sequences by Co(II) and Cu(II) ' PROTEIN SCIENCE , vol 16 , no. 8 , pp. 1751-1761 . DOI: 10.1110/ps.072846407 en
dc.identifier.issn 0961-8368
dc.identifier.other PURE UUID: 6d764719-1888-48ee-8a85-2cd89639b57d
dc.identifier.other PURE ITEMURL:
dc.identifier.other PURE LINK:
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dc.description.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. en
dc.format.extent 11
dc.format.extent 1751-1761
dc.format.mimetype application/pdf
dc.language.iso en en
dc.relation.ispartofseries PROTEIN SCIENCE en
dc.relation.ispartofseries Volume 16, issue 8 en
dc.rights openAccess en
dc.title Cleavage of recombinant proteins at poly-His sequences by Co(II) and Cu(II) en
dc.type A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä fi
dc.description.version Peer reviewed en
dc.contributor.department Department of Chemistry
dc.contributor.department VTT Technical Research Centre of Finland
dc.subject.keyword artificial protease
dc.subject.keyword Co(II)
dc.subject.keyword Cu(II)
dc.subject.keyword His tag
dc.subject.keyword protein cleavage
dc.subject.keyword protein stability
dc.identifier.urn URN:NBN:fi:aalto-201609234311
dc.identifier.doi 10.1110/ps.072846407
dc.type.version acceptedVersion

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