title: Sequence-specific Ni(II)-dependent peptide bond hydrolysis for protein engineering: active sequence optimization.
abstract: In previous studies we showed that Ni(II) ions can hydrolytically cleave a peptide bond preceding Ser/Thr in peptides of a general sequence RN-(Ser/Thr)-Xaa-His-Zaa-RC, where RN and RC are any peptide sequences. A peptide library screening, assisted by accurate measurements of reaction kinetics for selected peptides, demonstrated the preference for bulky and aromatic residues at variable positions Xaa and Zaa [A. Krężel, E. Kopera, A.M. Protas, A. Wysłouch-Cieszyńska, J. Poznański, W. Bal, J. Am. Chem. Soc., 132 (2010) 3355-3366]. In this work we used a similar strategy to find out whether the next residue downstream to Zaa may influence the reaction rate. Using an Ac-Gly-Ala-Ser-Arg-His-Zaa-Baa-Arg-Leu-NH2 library, with Zaa and Baa positions containing all common amino acids except of Cys, we found a very strong preference for aromatic residues in both variable positions. This finding significantly limits the range of useful Xaa, Zaa and Baa substitutions, thus facilitating the search for optimal sequences for protein engineering applications [E. Kopera, A. Belczyk-Ciesielska, W. Bal, PLoS One 7 (2012) e36350].
authors: Lodz Regional Park of Science and Technology Ltd., 114/116 Dubois Street, 93-465 Lodz, Poland.
authors: Anna Maria
authors: AM
authors: Protas
authors: Hanieh Hossein Nejad
authors: HH
authors: Ariani
authors: Arkadiusz
authors: A
authors: Bonna
authors: Agnieszka
authors: A
authors: Polkowska-Nowakowska
authors: Jarosław
authors: J
authors: Poznański
authors: Wojciech
authors: W
authors: Bal
mesh_terms: D002851:Chromatography, High Pressure Liquid; D006868:Hydrolysis; D008958:Models, Molecular; D009532:Nickel / Q000737:chemistry*; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry; D015202:Protein Engineering; D020539:Sequence Analysis, Protein; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D010455:Peptides; D009532:Nickel
keywords: Nickel(II) complex; Peptide bond hydrolysis; Peptide library; Rate constant