Letter: Mass spectrometric evidence for iron binding to the neuroprotective peptide NAP and its Cys5 mutant.

The NAP peptide (H(2)N-(1)NAPVSIPQ(8)-CONH(2)) is a truncated version of the activity-dependent neuroprotective protein. Its neuroprotective activities consist of the inhibition of Aß(25-35) and Aß(1-40) fibrillogenesis as well as protection against Aß-induced neurotoxicity and prevention of microtubule disruption associated with Alzheimer's disease. Therefore, we synthesized NAP and its mutant peptide with the sequence: H(2)N-(1)NAPVCIPQ(8)-COOH (NAPCOH), by replacing serine S(5) with cysteine C(5). Both native and mutant peptides were further used to study their interaction with iron ions. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry, Fourier transform infrared spectroscopy and also atomic force microscopy were used to probe Fe(3+) binding to both peptides. Contrary to the expected results, the investigated peptides underwent different oxidation processes, with resultant reduced Fe(2+) ions. These ions, and not the original Fe(3+) ions, were found to bind to each of non-oxidized peptides.