The N-terminal lobe fragment of hen ovotransferrin was prepared by trypsin digestion and the following gel filtration and ion-exchange chromatography. EPR spectra of Cu(II) complex of the N-terminal fragment were recorded independently from the C-terminal lobe and a major spectrum of the copper(II) complex showed g(//) = 2.300 and g(perpendicular to) = 2.061 with A(//)(Cu) = 14.0 mT, A(perpendicular to)(Cu) = 2.20 mT, which had a superhyperfine splitting (A(perpendicular to)(N) = 1.44 mT) of a nitrogen ligand, which may correspond to type A-1 site of copper binding position in the N-terminal lobe of hen ovotransferrin.