Application of Ni(II)-assisted peptide bond hydrolysis to non-enzymatic affinity tag removal.

PloS One
Edyta KoperaWojciech Bal

Abstract

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-His(6). 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.

References

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Citations

Apr 15, 2014·Journal of Inorganic Biochemistry·Ewa Izabela PodobasWojciech Bal
Oct 21, 2014·Chemical Research in Toxicology·Nina E WezynfeldTomasz Frączyk
Feb 23, 2013·Inorganic Chemistry·Hanieh H ArianiWojciech Bal
Jul 17, 2018·Metallomics : Integrated Biometal Science·Agnieszka Belczyk-CiesielskaWojciech Bal
Mar 30, 2019·Nature Methods·Bobo DangWilliam F DeGrado
Aug 17, 2020·Physical Review Letters·Ian L MorganOmar A Saleh
Aug 27, 2013·Journal of Inorganic Biochemistry·Anna Maria ProtasWojciech Bal

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