UV-Vis Spectroscopy Study on Interaction between Microperoxidase-11 and Pr Ion Under Acid Rain Stress

Interaction between rare earth ion praseodymium (Pr(Ⅲ)) and MP11 with/without hydrogen ion (H +) in different media( aqueous, phosphate buffer, physiological condition) were studied by UV Vis spectroscopy. All the results indicate that Pr(Ⅲ) interacts with MP11, increasing the non planarity of porphyrin periphery, leading MP11 to form two conformations when titrated by Pr(Ⅲ). Excessive Pr(Ⅲ) acts as a contaminant in living organism. H + and Pr(Ⅲ) have antagonistic effect on MP11, suggesting that at suitable concentration under physiological conditions, Pr(Ⅲ) can be used as biomodulator in protecting plants from acid rain stress or in rehabilitating the harm.

Rebinding kinetics of dissociated amino acid ligand and carbon monoxide to ferrous microperoxidase-11 in aqueous solution

The rebinding kinetics of an amino acid ligand to ferrous microperoxidase-11 (MP11) after photolysis of aggregated ferrous MP11 was measured in aqueous solution with femtosecond transient visible absorption spectroscopy. The kinetics of CO rebinding to ferrous MP11 after photolysis of MP11CO was also measured in aqueous solution with femtosecond transient visible absorption spectroscopy. From these measurements, we found that either Val-11 or Lys-13 rebinds to ferrous MP11 exponentially with an 8 picosecond time constant in aggregated ferrous MP11 solution and that CO rebinds to ferrous MP11 nonexponentially with subnanosecond time scale in MP11CO solution. The kinetics of both the amino acid and CO rebinding to ferrous MP11 in MP11 system mimics that in carbon monoxide oxidation activator protein (CooA) or carboxymethyl cytochrome c (CmCytC) system. We also measured the kinetics of CO rebinding to ferrous MP11 in aqueous solution at different MP11CO concentrations and found that MP11CO concentration has an obvious effect on the kinetics of CO rebinding to ferrous MP11, where both the germinate yield and rate of CO rebinding to ferrous MP11 increase with the increase of MP11CO concentration. These findings suggested that the picosecond amino acid ligand rebinding process could disturb the proximal heme-ligand structure that possibly leads to the subnanosecond CO rebinding kinetics in MP11CO, CooACO and CmCytCCO systems.

Screen, Design and Enzymatic Activity Determination of Artificial Microperoxidases

Peroxidase activity greatly impacts the maintenance of flee radical homeostasis, and can prevent or treat diseases related to free radicals. Microperoxidase-11（MP-11 ） is created via hydrolysis of cytochrome c iron-porphyrin complexes. In these complexes, the heme iron is penta-coordinate with histidine and exhibits excellent antioxidant activity when decomposing hydrogen peroxide. In this study, we screened the Ph.D.-7 and Ph.D.-12 phage display＇ peptide libraries and obtained ten small peptide ligands of deuterohemin（the vinyl groups of oxidized heme）. Among these polypeptides, DhHP-7P1, 12P1, 12P2 and 12P6 have good enzymatic activity compared with MP-11, and exhibit activities up to 50% of MP-11. Based on the screened sequences, we designed a series of artificial microperoxidases and determined that a higher peroxidase activity could be achieved with an enzymatic active site containing a second site of histidine residue spaced between two arginine residues with an interval of two amino acids（Dh-XHRXXR）.