Zinc finger peptide cleavage by a dinuclear platinum compound

Mangrum, John B.; Zgani, Ibrahim; Tsotsoros, Samantha D.; Qu, Yun; Farrell, Nicholas

doi:10.1039/c3cc44219e

Cited by 4 publications

(1 citation statement)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…49 Change of the protein structure was also supported in the presence of other metal ions, such as Cd(II) and Hg(II), 50 and the ZF cleavage was initiated by a dinuclear Pt(II) complex initially replacing the Zn(II) ions from the thiolate sites. 51 The structural integrity of P-1MEY and P-1MEY# proteins in the presence and absence of Ni(II) ions was verified by CD spectroscopy upon iterative heating and cooling (from 10 1C to 50 1C and back) of solutions adequate for the hydrolytic experiments. The collapse of the structure of ZF proteins at high temperatures would result in a disordered structure and thus, in an altered spectral pattern, similar to that obtained for the apo-protein (Fig.…”

Section: Monitoring the Ni(ii)-induced Hydrolysis Of The Zf Proteinsmentioning

confidence: 99%

Nickel(ii)-promoted specific hydrolysis of zinc finger proteins

et al. 2018

View full text Add to dashboard Cite

In this work we demonstrate that the previously described reaction of sequence specific Ni(ii)-dependent hydrolytic peptide bond cleavage can be performed in complex metalloprotein molecules, such as the Cys2His2 zinc finger proteins. The cleavage within a zinc finger unit possessing a (Ser/Thr)-X-His sequence is not hindered by the presence of the Zn(ii) ions. It results in loss of the Zn(ii) ion, oxidation of the SH groups and thus, in a collapse of the functional structure. We show that such natural Ni(ii)-cleavage sites in zinc finger domains can be edited out without compromising the DNA binding specificity. Inserting a Ni(ii)-susceptible sequence between the edited zinc finger and an affinity tag allows for removal of the latter sequence by Ni(ii) ions after the protein purification. We have shown that this reaction can be executed even when a metal ion binding N-terminal His-tag is present. The cleavage product maintains the native zinc finger structure involving Zn(ii) ions. Mass spectra revealed that a Ni(ii) ion remains coordinated to the hydrolyzed protein product through the N-terminal (Ser/Thr)-X-His tripeptide segment. The fact that the Ni(ii)-dependent protein hydrolysis is influenced by the Ni(ii) concentration, pH and temperature of the reaction provides a platform for novel regulated DNA effector design.

show abstract

Section: Monitoring the Ni(ii)-induced Hydrolysis Of The Zf Proteinsmentioning

confidence: 99%