Spontaneous deamidation in the
Asn-Gly-Arg (NGR) motif that yields an isoAsp-Gly-Arg (isoDGR) sequence has recently attracted considerable attention because of the possibility of application to dual
tumor targeting. It is well known that Asn deamidation reactions in
peptide chains occur via the five-membered ring
succinimide intermediate. Recently, we computationally showed by the B3LYP density functional theory method, that
inorganic phosphate and the Arg side chain can catalyze the NGR deamidation using a
cyclic peptide, c[CH₂CO⁻NGRC]⁻NH₂. In this previous study, the tetrahedral intermediate of the
succinimide formation was assumed to be readily protonated at the
nitrogen originating from the Asn side chain by the
solvent water before the release of an NH₃ molecule. In the present study, we found a new mechanism for the decomposition of the tetrahedral intermediate that does not require the protonation by an external
proton source. The computational method is the same as in the previous study. In the new mechanism, the release of an NH₃ molecule occurs after a
proton exchange between the
peptide and the
phosphate and conformational changes. The rate-determining step of the overall reaction course is the previously reported first step, i.e., the cyclization to form the tetrahedral intermediate.