Multiple roles of haem in cystathionine b-synthase activity: implications for hemin and other therapies of acute hepatic porphyria

The role of haem in the activity of cystathionine b-synthase (CBS) is reviewed and a hypothesis postulating multiple effects of haem on enzyme activity under conditions of haem excess or deficiency is proposed, with implications for some therapies of acute hepatic porphyrias. CBS utilises both haem and pyridoxal 5¢-phosphate (PLP) as cofactors. Although haem does not participate directly in the catalytic process, it is vital for PLP binding to the enzyme and potentially also for CBS stability. Haem deficiency can therefore undermine CBS activity by impairing PLP binding and facilitating CBS degradation. Excess haem can also impair CBS activity by inhibiting it via CO resulting from haem induction of haem oxygenase, and by induction of a functional vitamin B6 deficiency following activation of hepatic tryptophan 2,3-dioxygenase and subsequent utilisation of PLP by enhanced kynurenine aminotransferase and kynureninase activities. CBS inhibition results in accumulation of the cardiovascular risk factor homocysteine (Hcy) and evidence is emerging for plasma Hcy elevation in patients with acute hepatic porphyrias. Decreased CBS activity may also induce a proinflammatory state, inhibit expression of haem oxygenase and activate the extrahepatic kynurenine pathway thereby further contributing to the Hcy elevation. The hypothesis predicts likely changes in CBS activity and plasma Hcy levels in untreated hepatic porphyria patients and in those receiving hemin or certain gene-based therapies. In the present review, these aspects are discussed, means of testing the hypothesis in preclinical experimental settings and porphyric patients are suggested and potential nutritional and other therapies are proposed.

Exogenous hemin improves Cd2+ tolerance and remediation potential in Vigna radiata by intensifying the HO-1 mediated antioxidant defence system

The present study evaluated the effects of exogenous hemin on cadmium toxicity in terms of metal accretion and stress resilience in Vigna radiata L. (Wilczek). One-week-old seedlings were treated with CdCl2 (50 μM) alone and in combination with hemin (0.5 mM) in half-strength Hoagland medium for 96 h. The optimum concentrations of Cd and hemin were determined on the basis of haem oxygenase-1 activity. The results demonstrated that under Cd stress, plants accumulated a considerable amount of metal in their tissues, and the accumulation was higher in roots than in leaves, which significantly reduced the plant biomass and chlorophyll content by increasing the oxidative stress (MDA and H2O2 content). However, hemin supplementation under Cd,-stress improved plant growth by enhancing the harvestable biomass and photosynthetic pigments, increasing antioxidant activities (SOD, APX, POD, HO-1 and proline), lowering oxidative damage and increasing Cd tolerance in plants. Furthermore, the application of hemin enhances the removal efficiency of Cd in V. radiata by increasing the uptake of Cd via roots and its translocation from roots to foliar tissues. Thus, the study suggests that hemin has the potential to improve the stress tolerance and phytoremediation ability of heavy metal-tolerant plants so that they can be used instead of hyperaccumulators for remediation of Cd-contaminated environments.

A Dynamic Substrate is Required for MhuD-catalysed Degradation of Haem to Mycobilin

The non-canoncial haem oxygenase MhuD from <i>Mycobacterium tuberculosis</i> binds a haem substrate that adopts a dynamic equilibrium between planar and out-of-plane ruffled conformations. MhuD degrades this substrate to an unusual mycobilin product via successive monooxygenation and dioxygenation reactions. This article establishes a causal relationship between haem substrate dynamics and MhuD-catalysed haem degradation resulting in a revised enzymatic mechanism. UV/Vis absorption (Abs) and electrospray ionisation mass spectrometry (ESI-MS) data demonstrated that a second-sphere substitution favouring population of the ruffled haem conformation changed the rate-limiting step of the reaction resulting in a measurable build-up of the monooxygenated meso-hydroxyhaem intermediate. In addition, UV/Vis Abs and ESI-MS data for a second-sphere variant that favoured the planar substrate conformation showed that this change altered the enzymatic mechanism resulting in an alpha-biliverdin product. Single-turnover kinetic analyses for three MhuD variants revealed that the rate of haem monooxygenation depends upon the population of the ruffled substrate conformation. These kinetic analyses also revealed that the rate of meso-hydroxyhaem dioxygenation by MhuD depends upon the population of the planar substrate conformation. Thus, the ruffled haem conformation supports rapid haem monooxygenation by MhuD, but further oxygenation to the mycobilin product is inhibited. In contrast, the planar substrate conformation exhibits altered haem monooxygenation regiospecificity followed by rapid oxygenation of meso-hydroxyhaem. Altogether, these data yielded a revised enzymatic mechanism for MhuD where access to both substrate conformations is needed for rapid incorporation of three oxygen atoms into haem yielding mycobilin.<br>

Haem oxygenases play a pivotal role in placental physiology and pathology

BACKGROUND Haem oxygenases (HO) catabolise haem, which is the prosthetic group of numerous haemoproteins. Thus, multiple primary cellular pathways and functions rely on haem availability. HO exists in two isoforms, both expressed in the placenta, namely HO-1 and HO-2, the first being inducible. Haem oxygenases, particularly HO-1, have garnered specific interest in the field of physiological and pathological placental function. These enzymes mediate haem degradation by cleaving the alpha methene bridge to produce biliverdin, which is subsequently converted to bilirubin, carbon monoxide and iron. HO-1 has anti-inflammatory and antioxidant activities. SEARCH METHODS An initial literature analysis was performed using PubMed on 3 October 2018 using key terms such as ‘haem oxygenase and pregnancy’, ‘haem oxygenase and placenta’, ‘HO-1 and pregnancy’, ‘HO-1 and placenta’, ‘HO and placenta’, ‘HO and pregnancy’, ‘genetic variant and HO’, ‘CO and pregnancy’, ‘CO and placenta’, ‘Bilirubin and pregnancy’, ‘Iron and pregnancy’ and ‘PPAR and Haem’, selecting consensus conferences, recommendations, meta-analyses, practical recommendations and reviews. A second literature analysis was performed, including notable miscarriages, foetal loss and diabetes mellitus, on 20 December 2019. The three authors studied the publications independently to decipher whether they should be included in the manuscript. OBJECTIVE AND RATIONALE This review aimed to summarise current pieces of knowledge of haem oxygenase location, function and regulation in the placenta, either in healthy pregnancies or those associated with miscarriages and foetal loss, pre-eclampsia, foetal growth restriction and diabetes mellitus. OUTCOMES HO-1 exerts some protective effects on the placentation, probably by a combination of factors, including its interrelation with the PGC-1α/PPAR pathway and the sFlt1/PlGF balance, and through its primary metabolites, notably carbon monoxide and bilirubin. Its protective role has been highlighted in numerous pregnancy conditions, including pre-eclampsia, foetal growth restriction, gestational diabetes mellitus and miscarriages. WIDER IMPLICATIONS HO-1 is a crucial enzyme in physiological and pathological placentation. This protective enzyme is currently considered a potential therapeutic target in various pregnancy diseases.

Carbon monoxide in intensive care medicine—time to start the therapeutic application?!

Carbon monoxide (CO) is not only known as a toxic gas due to its characteristics as an odorless molecule and its rapid binding to haem-containing molecules, thus inhibiting the respiratory chain in cells resulting in hypoxia. For decades, scientists established evidence about its endogenously production in the breakdown of haem via haem-oxygenase (HO-1) and its physiological effects. Among these, the modulation of various systems inside the body are well described (e.g., anti-inflammatory, anti-oxidative, anti-apoptotic, and anti-proliferative). Carbon monoxide is able to modulate several extra- and intra-cellular signaling molecules leading to differentiated response according to the specific stimulus. With our growing understanding in the way CO exerts its effects, especially in the mitochondria and its intracellular pathways, it is tempting to speculate about a clinical application of this substance. Since HO-1 is not easy to induce, research focused on the application of the gaseous molecule CO by itself or the implementation of carbon monoxide releasing molecules (CO-RM) to deliver the molecule at a time- and dose dependently safe way to any target organ. After years of research in cellular systems and animal models, summing up data about safety issues as well as possible target to treat in various diseases, the first feasibility trials in humans were established. Up-to-date, safety issues have been cleared for low-dose carbon monoxide inhalation (up to 500 ppm), while there is no clinical data regarding the injection or intake of any kind of CO-RM so far. Current models of human research include sepsis, acute lung injury, and acute respiratory distress syndrome as well as acute kidney injury. Carbon monoxide is a most promising candidate in terms of a therapeutic agent to improve outbalanced organ conditions. In this paper, we summarized the current understanding of carbon monoxide’s biology and its possible organ targets to treating the critically ill patients in tomorrow’s ICU.