Elsevier

Nitric Oxide

Volume 87, 1 June 2019, Pages 83-89
Nitric Oxide

The origins of nitric oxide and peroxynitrite research in Uruguay: 25 years of contributions to the biochemical and biomedical sciences

https://doi.org/10.1016/j.niox.2019.03.003Get rights and content

Highlights

  • •

    A historical background on how nitric oxide (radical dotNO) and peroxynitrite research originated in Uruguay is provided.

  • •

    •NO shifts its signal transduction action towards oxidative processes secondary to its reaction with superoxide radical (O2radical dot–).

  • •

    The diffusion-controlled reaction of radical dotNO with O2radical dot yields peroxynitrite, a strong biological oxidant

  • •

    The evolution of the biochemistry and cell biology of peroxynitrite is presented.

  • •

    Hypothesis and mechanistic-driven research in radical dotNO and peroxynitrite serves to understand and treat medically-relevant conditions.

Abstract

In this review, I provide historical background on how nitric oxide (•NO) and peroxynitrite research was originated in Uruguay in the early 90′s and how the investigations evolved through over more than two decades within a context related to human biology. This process involved the participation of multiple local investigators, in conjunction with collaborations at the regional (Latin American) and international levels. The discoveries have been integrated with parallel investigations from other research groups worldwide and, have provided a body of knowledge to unravel how the free radical nitric oxide (•NO) can shift its signal transduction action towards oxidative processes via its interactions with superoxide radical (O2•–) to yield peroxynitrite, a strong biological oxidant. The oxidative biochemistry of peroxynitrite involves both direct reactions and the formation of secondary oxidizing species (i.e. hydroxyl radicals (•OH), carbonate radicals (CO3•–) and nitrogen dioxide (•NO2)) that cause oxidative modifications of biomolecules, including thiol oxidation and tyrosine nitration. Due to the intrinsic instability of peroxynitrite in biological systems, its half-life and fate are largely dictated by its reaction kinetics with biotargets. The direct actions of •NO and peroxynitrite in the modulation of intracellular redox processes are disparate, with peroxynitrite typically causing permanent modifications of cellular components and resulting in severe alterations of cell and mitochondrial homeostasis. Herein, I highlight the evolution and progression of •NO and peroxynitrite research in Uruguay during over 25 years of work, emphasizing hypothesis- and mechanistic-oriented biochemical studies and their translation to medically-relevant conditions.

Introduction

This review, part of a special issue dedicated on science in Latin America, provides historical background on how nitric oxide (•NO) and peroxynitrite research was originated in Uruguay in the early 90′s and how the investigations evolved through over more than two decades with a strong biochemical basis and translated to the biomedical sciences. Due to the nature of this article, the reader will be referred to significant number of works from our group and integrated with other parallel contributions. I will particularly quote collaborative research activities that we have performed with other Latin American groups mainly based in Argentina, Chile and Brazil and that have been essential to keep a strong research program. Overall, the article highlights a discovery process based at our laboratories at the Universidad de la República in Montevideo, Uruguay, that contributed to create a solid body of knowledge to unravel at the molecular level how the free radical •NO shifts its signal transduction action towards oxidative processes via its interactions with superoxide radical (O2•–) to yield peroxynitrite, a strong biological oxidant. In parallel to the main specific discoveries described herein, the research program implicated the simultaneous training of various generations of local and regional scientists and the development of research infrastructures.

Section snippets

The tradition of free radical and redox research in Uruguay

In the mid 80's, I started to perform work on xanthine oxidase enzymology characterizing O2•– formation by chemiluminescence probes [1,2]. This type of studies in our Department were, in fact, initiated in the late 50's during a sabbatical visit of Dr. John R. Totter [3,4], an (north) American biochemist, and continued in the 60's and early 70's by one of his disciples, Dr. Eugenio Prodanov [5,6]. This advanced research program was halted in the early 70's due to the irruption of a military

The incorporation of nitric oxide and peroxynitrite research in Uruguay

Upon my return back to Uruguay in 1992, I was committed to continue with •NO and peroxynitrite research, as during my three years at UAB, we made a number of observations that clearly showed us that this was potentially a groundbreaking area, and that the only way to consolidate the “peroxynitrite theory” was to keep up with contributions and experiments for the next several years (Fig. 1). The initial years of “peroxynitrite in biology” were seen with some skepticism by many researchers, in

Modulatory action of nitric oxide on lipid peroxidation and the formation of nitrolipids

In 1994, Bruce A. Freeman visited our laboratories in Montevideo under the auspices of the Fulbright Foundation. After publishing together at UAB the observation that peroxynitrite could promote lipid peroxidation [13] and an independent observation indicating that simultaneous fluxes of •NO and O2•– (the precursors of peroxynitrite) led to human low density lipoprotein lipid peroxidation [38], Bruce and us became interested to investigate whether lipid peroxidation yields and product

Peroxynitrite in neurodegeneration

In the mid 90′s Luis Barbeito, a fine Uruguayan neurobiologist (at the time located at the Instituto de Investigaciones Biológicas Clemente Estable and currently at the Pasteur Institute of Montevideo) became very enthusiastic to initiate work together on the role of •NO, oxidative stress and peroxynitrite in neurodegenerative processes. At the time I contacted Joe S. Beckman (that had moved from UAB to Oregon State University, OSU) to join our research plans as he was already interested on the

Studies on protein tyrosine nitration

The capacity of peroxynitrite to cause the nitration of protein tyrosine residues was observed in the early 90′s at UAB by Ischiropoulos, Beckman et al. [15,16]. Tyrosine nitration was initially conceived as an oxidative posttranslational modification and a “footprint” of the reactions of peroxynitrite, a concept that was later extended to other •NO-derived oxidants (reviewed in Refs. [14,52]). A large number of contributions have shown the presence of tyrosine nitrated proteins in human

Biological half-life of peroxynitrite and permeation across biomembranes

In the 90's a large amount of kinetic data was gathered (including a growing list of second order rate constants of peroxynitrite with biomolecules) with the use of both direct stopped flow spectrophotometry following peroxynitrite decay at 302 nm and by competition kinetics [72]. Then, it became evident that peroxynitrite would be readily consumed in biological systems and with an estimated half-life in the ms range. Considering a diffusion constant for peroxynitrite similar to that of nitrate

Peroxynitrite detoxification systems in pathogens in virulence

The cytotoxic action of peroxynitrite against invading microorganisms is modulated by the endogenous antioxidant armamentarium of the pathogens [80]. In this regard, the antioxidant enzyme networks of pathogenic microorganisms are now considered to participate in their virulence to host cells, including macrophages. Notably, a family of thiol-containing enzymes, the peroxiredoxins have been shown to participate in the catalytic two-electron reduction of peroxynitrite to nitrite and attenuate

Peroxynitrite in the vasculature and human research studies

The interactions of endothelial-derived •NO with vascular derived O2•– were appreciated early on [90,91] and excess O2•– leads to decreased bioavailability of •NO and endothelial dysfunction. In addition, there is a consequent formation of peroxynitrite that can participate in vascular damage during degenerative processes related to, for example, atherosclerosis, hyperglycemia and hypertension and aging [17,92]. Our group has contributed to understand the molecular basis of enhanced O2•–

Redox-based therapeutics to neutralize peroxynitrite

In the mid to late 90′s we initiated work in collaboration with Ines Batinic-Haberle, Irwin Fridovich and collaborators at Duke University to assess the reactions of Mn-porphyrins (MnP) with peroxynitrite. At the time MnP were seen as “SOD mimics” [96] but it readily became apparent that the redox and cytoprotective actions that MnP exerted in cellular systems and even in vivo, were likely due to redox mechanisms other than simply O2•- dismutation, due to kinetic considerations. In this

Concluding remarks

A research journey on •NO and peroxynitrite research started in Uruguay over 25 years ago. While of strong biochemical basis, the studies evolved through more than two decades encompassing several disciplines ranging from physical-chemistry to translational medicine. The body of accumulated research allowed to create and solidify a hypothesis that links •NO with redox metabolism in relation to human disease conditions and the process of aging. This research flourished in Uruguay with now

Acknowledgements

This work was supported by current grants from Universidad de la República (CSIC Grupos 2014, Espacio Interdiciplinario 2015) and Agencia Nacional de Investigación e Innovación (FCE_1_2014_1_104233). Additional support was obtained through Fundación Manuél Pérez and Programa de Desarrollo de Ciencias Básicas (PEDECIBA), Uruguay. I also gratefully acknowledge the contribution of other research funding agencies at various different times including support from the Howard Hughes Medical Institute,

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