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Updated: Jul 25, 2025

Measurement of Heme Synthesis Levels in Mammalian Cells
Published on: July 9, 2015
Heme protonation affects iron-NO binding in the NO transport protein nitrophorin
Hendrik Auerbach1, Isabelle Faus1, Sergej Rackwitz1
1Department of Physics, RPTU Kaiserslautern-Landau, 67663 Kaiserslautern, Germany.
Abstract:
The nitrophorins (NPs) comprise an unusual group of heme proteins with stable ferric heme iron nitric oxide (Fe-NO) complexes. They are found in the salivary glands of the blood-sucking kissing bug Rhodnius prolixus, which uses the NPs to transport the highly reactive signaling molecule NO. Nuclear resonance vibrational spectroscopy (NRVS) of both isoform NP2 and a mutant NP2(Leu132Val) show, after addition of NO, a strong structured vibrational band at around 600 cm-1, which is due to modes with significant Fe-NO bending and stretching contribution. Based on a hybrid calculation method, which uses density functional theory and molecular mechanics, it is demonstrated that protonation of the heme carboxyl groups does influence both the vibrational properties of the Fe-NO entity and its electronic ground state. Moreover, heme protonation causes a significant increase of the gap between the highest occupied and lowest unoccupied molecular orbital by almost one order of magnitude leading to a stabilization of the Fe-NO bond.
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