Adrenaline by ferrylmyoglobin

Cecilia R Giulivi, Enrique Cadenas

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The oxidation of adrenaline by ferrylmyoglobin, the product formed by the oxidation of myoglobin with H2O2, was examined by absorption, fluorescence, and EPR spectroscopy in terms of the formation of intermediate free radicals and stable molecular products and the binding of adrenaline oxidation products to the apoprotein. The reaction of adrenaline with ferrylmyoglobin resulted in reduction of the hemoprotein to metmyoglobin and consumption of adrenaline. Quantification of metmyoglobin formed per adrenaline yielded a ratio of 1.66. The reaction was found first order on adrenaline concentration and second order on ferrylmyoglobin concentration. This, together with the above ratio, suggested a mechanism by which two oxoferryl moieties (ferrylmyoglobin) were reduced by adrenaline yielding metmyoglobin and the o-semiquinone state of adrenaline. The decay of the o- semiquinone to adrenochrome was confirmed by an increase in absorbance at 485 nm. The product was nonfluorescent; alkalinization of the reaction mixture resulted in a strong fluorescence at 540 nm ascribed to 3,5,6-trihydroxyindol or adrenolutin. Hence, adrenochrome and its alkali-catalyzed product, adrenolutin, are the major molecular products formed during the oxidation of adrenaline by ferrylmyoglobin. Semiquinones formed during the adrenaline/ferrylmyoglobin interaction were detected by EPR, spin stabilizing these species with Mg2+. The six-line EPR spectrum observed (a(N) = 4.5 G, a(N)(CH3) = 5.1, and a(2H) = 0.91; g = 2.0040) may be assigned to the semiquinone forms of adrenochrome and/or adrenolutin or a composite of these species. The intensity of the EPR signal increased with time and its subsequent decay followed a second-order kinetics as inferred by the proportionality of the square of the EPR line intensity with H2O2 concentration. Heine destruction and lysine loss, inherent in the reaction of metmyoglobin with H2O2, were prevented 80 and 34% by adrenaline, respectively. The low protection exerted by adrenaline against lysine loss was possibly due to the formation of Schiff bases between the ε-NH2 group of lysine and the o-quinone oxidation product(s) of adrenaline. The yield of Schiff base formation was 20-25%. The autoxidation of adrenaline at physiological pH is extremely slow or nonexistent. These data provide a rationale for the primary oxidation of adrenaline by the pseudoperoxidatic activity of ferrylmyoglobin and suggest implications of the free radicals thereby formed for the oxidative damage in reperfusion injury.

Original languageEnglish (US)
Pages (from-to)175-183
Number of pages9
JournalFree Radical Biology and Medicine
Volume25
Issue number2
DOIs
StatePublished - Jul 15 1998
Externally publishedYes

Fingerprint

Epinephrine
Metmyoglobin
Adrenochrome
Paramagnetic resonance
Oxidation
Lysine
Schiff Bases
ferrylmyoglobin
Free Radicals
Fluorescence
Apoproteins
Myoglobin
Fluorescence Spectrometry
Alkalies
Reperfusion Injury
Spectroscopy

Keywords

  • Adrenaline
  • Ferrylmyoglobin
  • Free radicals
  • Ischemia/reperfusion
  • Myoglobin
  • Ortho-Semiquinones

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Medicine(all)
  • Toxicology

Cite this

Adrenaline by ferrylmyoglobin. / Giulivi, Cecilia R; Cadenas, Enrique.

In: Free Radical Biology and Medicine, Vol. 25, No. 2, 15.07.1998, p. 175-183.

Research output: Contribution to journalArticle

Giulivi, Cecilia R ; Cadenas, Enrique. / Adrenaline by ferrylmyoglobin. In: Free Radical Biology and Medicine. 1998 ; Vol. 25, No. 2. pp. 175-183.
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