C2+ Inhibits The Peroxidase And Antibacterial Activity Of Homodimer Hemoglobin From Blood Clam Tegillarca Granosa By Destroying Its Heme Pocket Structure

Beyond its role as an oxygen transport protein, the homodimer hemoglobin of blood clam Tegillarca granosa (Tg-Hbl) has been found to possess antibacterial activity. However, the mechanism of antibacterial activity of Tg-Hbl remain to be investigated. In this study, we investigated the effects of Cu2+ on the structure, peroxidase activity, and antibacterial ability of Tg-Hbl. Tg-Hbl was significantly inactivated by Cu2+ in a non-competitive inhibition manner, following first-order reaction kinetics. The Spectroscopy results showed that Cu2+ changed the iron porphyrin ring and the coordination of heme with proximal histidine of Tg-Hbl, and increased the hydrophobicity of heme pocket. We found that proline could stabilize the heme pocket structure of Tg-Hbl, hence, protect peroxidase activity and antimicrobial activity of Tg-Hbl against damage by Cu2+. Our results suggest that Cu2+ inhibits the peroxidase and antibacterial activity of Tg-Hbl by destroying its heme pocket structure and Tg-Hbl probably plays an antibacterial role through its peroxidase activity. This result could provide insights into the antibacterial mechanism of Tg-Hbl.