Dugad L B, Goff H M, Abeles F B
Department of Chemistry, University of Iowa, Iowa City 52242.
Biochim Biophys Acta. 1991 Dec 11;1118(1):36-40. doi: 10.1016/0167-4838(91)90438-6.
Two peroxidase isoenzymes from Cucumber seedlings, one acidic (pI = 4) and one basic (pI = 9), were characterized by 1H-NMR spectroscopy. The NMR spectra were obtained in the native (ferric high-spin) and cyanide ligated (ferric low-spin) forms of both isoenzymes. The NMR spectral comparison of paramagnetically shifted resonances with those of the well characterized horseradish peroxidase C, HRP(C), isoenzyme indicates that both cucumber peroxidases have a protohemin IX prosthetic group with proximal histidine coordinated to the heme iron. The downfield heme 1H-NMR shift pattern is distinct for each isoenzyme, and this reflects presumably dissimilar heme active site environments. The basic isoenzyme shows less asymmetry in heme 1H-NMR signals as compared to the acidic isoenzyme or HRP(C) isoenzyme. It was also found that the acidic cucumber peroxidase exists predominantly as a monomeric species in solution with 30 kDa molecular mass as opposed to its earlier characterization as a 60 kDa dimeric protein.
