A 1 H‐NMR study of electronic structure of the active site of Galeorhinus japonicus metmyoglobin

• Published in: European Journal of Biochemistry
• Main Authors: YAMAMOTO, Yasuhiko, ŌSAWA, Akemi, INOUE, Yoshio, CHÛJÔ, Riichirô, SUZUKI, Tomohiko
• Format: Article in Journal/Newspaper
• Language: English
• Published: Wiley 1990
• Subjects: Wyman; Sperm whale
• Online Access: http://dx.doi.org/10.1111/j.1432-1033.1990.tb19219.x; https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fj.1432-1033.1990.tb19219.x; https://febs.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1432-1033.1990.tb19219.x

The ferric high‐spin form of the myoglobin from the shark Galeorhinus japonicus , which possesses a Gln residue at the distal site instead of the usual His residue, has been studied by 1 H‐NMR spectroscopy. Using the heme meso ‐proton (C5H, C10H, C15H and C20H) resonance shift as a diagnostic probe for identifying the coordination system of the iron center in ferric high‐spin form of hemoprotein, it has been shown that G. japonicus metmyoglobin (metMb) possesses the pentacoordinated active site. The pH‐dependence study of NMR spectra of G. japonicus metMb revealed the appearance of the hydroxyl form of metMb at high pH, indicating that the protein undergoes the transition between the acidic and alkaline forms. The p K value and the rate for this acid‐alkaline transition in G. japonicus metMb were found to be ∼ 10 and ≪ 4 × 10 2 s −1 , respectively. Since the p K value of the acidalkaline transition for the pentacoordinated heme in Aplysia limacina metMb is 7.8 [Giacometti, G. M., Das Ros, A., Antonini, E. & Brunori, M. (1975) Biochemistry 14 , 1584–1588] and that of the hexacoordinated heme in sperm whale metMb is 9.1 [Brunori, M., Antonini, E., Fasella, P., Wyman, J. & Rossi‐Fanelli, A. (1968) J. Mol. Biol. 34 , 497.–504], the OH − affinity of the ferric heme iron does not appear to depend on its coordination system. The acid‐alkaline transition rate in A. limacina metMb was reported to be ≪ 1.5 × 10 2 s −1 [Pande, U., La Mar, G. N., Lecomte, J. T. J., Ascoli, F., Brunori, M., Smith, K. M., Pandey, R. K., Parish, D. W. & Thanabal, V. (1986) Biochemistry 25 , 5638–5646] and therefore a slow transition rate may be unique to the pentacoordinated active site of Mb.