Research Article | OPEN ACCESS
Spectroscopic Investigation of Pyruvate Formate Lyase-activating Enzyme: A Look into EPR, ENDOR and Mossabuer Spectroscopy
1Danilo O. Ortillo and 2Joan B. Broderick
1Department of Chemistry, University of the Philippines Visayas, 5023 Miagao, Iloilo, Philippines
2Department of Chemistry and Biochemistry, 103 Chemistry and Biochemistry Building, P.O. Box 173400, Bozeman, MT 59717, USA
Research Journal of Applied Sciences, Engineering and Technology 2014 9:1075-1097
Received: April ‎08, ‎2014 | Accepted: April ‎28, ‎2014 | Published: September 05, 2014
Abstract
Electron Paramagnetic Resonance (EPR) and Electron Nuclear Double Resonance (ENDOR) spectroscopies are extremely powerful and versatile methods for the characterization of paramagnetic systems in biology, chemistry and physics. For iron centers in the radical SAM enzymes however, Mössbauer spectroscopy has proven to be both powerful and useful as a complementary spectroscopic technique in determining not just the oxidation states but also the type of iron species present in the catalytic center. The cluster content of the radical SAM protein, Pyruvate Formate-Lyase-Activating Enzyme (PFL-AE), was characterized using EPR and Mössbauer techniques while additional ENDOR analysis helped determine the novel interaction of the co-substrate, S- Adenosylmethionine (SAM or AdoMet) with the Fe-S cluster of PFL-AE. The anchoring role of the Fe-S cluster to the co-substrate derived from the spectroscopic data supports the mechanism where a SAM-based radical species is involved during catalysis.
Keywords:
AdoMet , pyruvate formate-lyase-activating enzyme , radical SAM , spectroscopic methods,
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The authors have no competing interests.
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