Object's details: Application of Ni(II)-Assisted Peptide Bond Hydrolysis to Non-Enzymatic Affinity Tag Removal

Provider:Repozytorium Instytutu Biochemii i Biofizyki Polskiej Akademii Nauk

Description

Title:
Application of Ni(II)-Assisted Peptide Bond Hydrolysis to Non-Enzymatic Affinity Tag Removal
Creator:
Kopera, Edyta;
Belczyk-Ciesielska, Agnieszka;
Bal, Wojciech
Description:
In this study, we demonstrate a non-enzymatic method for hydrolytic peptide bond cleavage, applied to the removal of an affinity tag from a recombinant fusion protein, SPI2-SRHWAP-His6. This method is based on a highly specific Ni(II) reaction with (S/T)XHZ peptide sequences. It can be applied for the protein attached to an affinity column or to the unbound protein in solution. We studied the effect of pH, temperature and Ni(II) concentration on the efficacy of cleavage and developed an analytical protocol, which provides active protein with a 90% yield and ,100% purity. The method works well in the presence of non-ionic detergents, DTT and GuHCl, therefore providing a viable alternative for currently used techniques.
Object availability:
free access
Date:
2012.05.04
Type:
Article;
PeerReviewed
Language:
English
Format:
application/pdf
Relation: http://eprints.ibb.waw.pl/511 ;http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0036350 ;
10.1371/journal.pone.0036350
Subject:
Q Science (General)
Identifier: http://eprints.ibb.waw.pl/511/1/PLoSOne_SPI2hydrolysis_2012.pdf ;
Kopera, Edyta and Belczyk-Ciesielska, Agnieszka and Bal, Wojciech (2012) Application of Ni(II)-Assisted Peptide Bond Hydrolysis to Non-Enzymatic Affinity Tag Removal. PloS one, 7 (5). pp. 1-9. ISSN 1932-6203
Data provider:
Repozytorium Instytutu Biochemii i Biofizyki Polskiej Akademii Nauk

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Object's details: Application of Ni(II)-Assisted Peptide Bond Hydrolysis to Non-Enzymatic Affinity Tag Removal

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