Heme active-site structural characterization of chloroperoxidase by resonance Raman spectroscopy.
Resonance Raman spectra are reported for the nitric oxide adducts of ferric and ferrous chloroperoxidase and carbon monoxide adducts of ferrous chloroperoxidase. The stretching, v(Fe-NO), and bending, delta(FeNO), modes are detected at 538 and 558 cm-1, respectively, for the ferric nitrosylchloroperoxidase. These two bands shift to 534 and 546 cm-1, respectively, upon substitution by 15N16O. The v(Fe-NO) mode of the nitric oxide adduct of ferrous chloroperoxidase is located at 542 cm-1, which shifts to 528 (15N16O), 540 (14N18O), and 524 (15N18O) cm-1 as the mass of the bound nitric oxide increases by 1 atomic unit. Two distinct states of the carbon monoxide adduct of chloroperoxidase, the acidic and alkaline forms, are found to undergo a reversible pH-induced transition. The v(Fe-CO) mode shifts from 484 to 492 cm-1 and the delta(FeCO) mode at 562 cm-1 disappears as the pH is reduced from 6.0 to 3.3. In addition, two low frequency modes at 382 and 420 cm-1, assignable to the delta(CbC1C2) bending of propionate and vinyl groups, respectively, also show pH sensitivity. The results suggest a peroxidase-like heme active-site environment for chloroperoxidase and indicate a facile conformational change of heme groups accompanying the acid-base transition.
Chemistry Department, Marquette University, Milwaukee, Wisconsin 53233.
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Hu
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Kincaid
D001665:Binding Sites; D002711:Chloride Peroxidase / Q000737:chemistry / Q000378:metabolism*; D003545:Cysteine / Q000378:metabolism; D004563:Electrochemistry; D006418:Heme / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D009195:Peroxidase / Q000737:chemistry; D011487:Protein Conformation; D013059:Spectrum Analysis, Raman
D006418:Heme; D002711:Chloride Peroxidase; D009195:Peroxidase; D003545:Cysteine