Hemoglobin induction in mouse macrophages

^{Howard Hughes Medical Institute and}^{Department of Medicine, Pulmonary and Cardiology Divisions, Duke University Medical Center, Durham, NC 27710}

^{To whom reprint requests should be addressed at: MSRB Room 321 (Box 2612), Duke University Medical Center, Durham, NC 27710. e-mail:}ude.ekud.cm@100LMATS.

Communicated by Irwin Fridovich, Duke University Medical Center, Durham NC

Received 1999 Mar 3; Accepted 1999 Apr 20.

Abstract

The common perception that hemoglobin is involved solely in the transport of oxygen and carbon dioxide has been challenged by recent studies with nitric oxide (NO). These studies have shown that the primordial bacterial flavohemoglobin functions to consume NO enzymatically (to protect from nitrosative stress), whereas mammalian hemoglobin functions to deliver NO (thus maximizing oxygen delivery in the respiratory cycle). Here we report that murine macrophages stimulated to produce NO with lipopolysaccharide and interferon-γ express the β^{hemoglobin subunit. Consumption of NO, however, was not increased by cytokines or by hemoglobin expression. These data suggest alternative functions for globins in mammalian cells, and they challenge the prevailing view that the expression of α- and β-globin genes is always balanced and coordinated.}

Abstract

The common perception of hemoglobin is that it is selectively expressed in cells of erythroid lineage and involved solely in the transport of oxygen from the lungs to the tissues and of carbon dioxide from the tissues to the lungs. The function of hemoglobin has been interpreted in terms of an equilibrium between two alternative structures, one with high affinity for oxygen and the other with low affinity (1). Although the position of this equilibrium is governed by the spin state of the heme iron, it is a structural change in the heme pocket rather than a change in oxidation state of the iron that is thought to trigger deoxygenation (1). Thus, hemoglobins have been functionally distinguished from the heme-containing cytochromes, which carry out redox reactions with their ligands (2).

These general principles have been challenged by recent studies with nitric oxide (NO). First, hemoglobin transports not only oxygen but also NO, which it releases in tissues to dilate blood vessels (3, 4). Second, the binding and release of NO involves redox chemistry (5), blurring the functional distinction between hemoglobins and cytochromes. Third, it has been shown recently that a bacterial flavohemoglobin consumes NO enzymatically (6, 7). NO increases both oxygen consumption and NAD(P)H utilization by the flavohemoglobin (6), proving that it functions as an oxygenase that metabolizes NO (6), rather than as an oxidase that generates NO-scavenging superoxide (8). In addition, the flavohemoglobin reduces NO to N₂O anaerobically (6, 9). Thus, the primordial function of hemoglobin—a protein found not only in erythrocytes but also in microorganisms, invertebrates, and plants—may well be to protect against nitrosative stress (6, 7, 10).

In this new light, we questioned the remaining tenet that mammalian globin genes are normally expressed only in erythroid cell lineages. Cytokine-activated murine macrophages are a well-studied model of NO production (11–16). Specifically, treatment with cytokines leads to induction of NO synthase, activation of the NADPH oxidase, and inhibition of cell respiration. The mechanism by which these cells survive the high-level production of reactive oxygen and nitrogen species is not well understood. Here we show that treatment with lipopolysaccharide (LPS) and interferon (IFN)-γ leads to activation of the β^{-globin gene. It is likely, however, that hemoglobin carries out different functions in macrophages than in erythrocytes or bacteria.}

Acknowledgments

We thank Russel Kaufman for helpful comments and Alfred Hausladen for help with NO assays.

Acknowledgments

ABBREVIATIONS

IFN	interferon
LPS	lipopolysaccharide
RT-PCR	reverse transcription–PCR
GAPDH	glyceraldehyde-3-phosphate dehydrogenase

ABBREVIATIONS

Footnotes

Data deposition: The sequence reported in this paper has been deposited in the GenBank database (accession no. {"type":"entrez-nucleotide","attrs":{"text":"AF149782","term_id":"5305804"}}AF149782).

Footnotes

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