Vol 54(2020) N 6 p. 904-910; DOI 10.1134/S0026893320060035
P.V. Ershov1, Yu.V. Mezentsev1, E.O. Yablokov1, L.A. Kaluzgskiy1, A.S. Ivanov1, N.V. Gnuchev2, V.A. Mitkevich2, A.A. Makarov2, S.A. Kozin2*
Direct Molecular Fishing of Zinc-Dependent Protein Partners of Amyloid-beta 1-16 with the Taiwan (D7H) Mutation and Phosphorylated Ser8 Residue1Orekhovich Institute of Biomedical Chemistry, Moscow, 119121 Russia
2Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia
Received - 2020-06-11; Revised - 2020-07-10; Accepted - 2020-07-10
We previously showed that the metal-binding domain 1-16 of intact amyloid-beta (Aβ) is involved in interactions with a number of proteins from the cytosolic fraction of SK-N-SH human neuroblastoma cells in a zinc-dependent manner only. It is known that hereditary mutations in the Aβ metal-binding domain (Aβ(1-16)), which accelerate the development of Alzheimer's disease and post-translational modifications of amino acid residues, can significantly affect the domain's structure in the presence of zinc ions. In this work, using the molecular fishing methodology for Aβ(l-16) isoforms with the Taiwanese mutation (D7H) and a phosphorylated Ser8 residue, proteins from the cytosol of SK-N-SH cells were found that are able to form zinc-dependent non-covalent complexes with these domains. The partner proteins identified for these isoforms differed from those for intact Aβ(1-16). In contrast, the Aβ(1-16) isoform with the English mutation (H6R) and the Aβ(1-16) isoform containing both an isomerized Asp7 residue and phosphorylated Ser8 residue did not interact with cytosolic proteins. The results are useful for developing methods for rational modulation of protein-protein interactions involving natural isoforms of beta-amyloid, and also indicate the possible role of beta-amyloid with phosphorylated Ser8 as a molecule involved in normal physiological processes.
Alzheimer's disease, amyloid-beta, isoforms, metal-binding domain, zinc, protein-protein interactions, SK-N-SH cells