Nitrite binding to metmyoglobin and methemoglobin in comparison to nitric oxide binding

• Published in: Journal of biological inorganic chemistry 2002-01, Vol.7 (1-2), p.165-176
• Main Authors: Wanat, Alicja, Gdula-Argasińska, Joanna, Rutkowska-Zbik, Dorota, Witko, Małgorzata, Stochel, Grazyna, van Eldik, Rudi
• Format: Article
• Language: English
• Subjects: Animals; Hemoglobins - metabolism; Kinetics; Methemoglobin - metabolism; Metmyoglobin - metabolism; Nitric Oxide - metabolism; Nitrites - metabolism; Protein Binding - physiology; Thermodynamics
• ISSN: 0949-8257; 1432-1327
• DOI: 10.1007/s007750100284

Nitrite binds reversibly to the ferriheme proteins metmyoglobin and methemoglobin in aqueous buffer solution at a physiological pH of 7.4. The spectral changes recorded for the formation of metMb(NO2-) differ significantly from those observed for the nitrosylation of metMb, which can be accounted for in terms of the different reaction products. Nitric oxide binding to metMb produces a nitrosyl product with Fe(II)-NO+ character, whereas the reaction with nitrite produces an Fe(III)-NO2- complex. The kinetics of the binding and release of nitrite by metMb and metHb were investigated by stopped-flow techniques at ambient and high pressure. The kinetic traces recorded for the reaction of nitrite with metMb exhibit excellent single-exponential fits, whereas nitrite binding to metHb is characterized by double-exponential kinetics which were assigned to the reactions of the alpha- and beta-chains of metHb with NO2-. The rate constants for the binding of nitrite to metMb and metHb were found to be much smaller than those reported for the binding of NO, such that nitrite impurities will not affect the latter reaction. The activation parameters (deltaH++,deltaS(ne),deltaV++) obtained from the temperature and pressure dependence of the reactions support the operation of a dissociative mechanism for the binding and release of nitrite, similar to that found for the binding and release of NO in metMb.