title: Novel insights in mammalian catalase heme maturation: effect of NO and thioredoxin-1.
abstract: Catalase is a tetrameric heme-containing enzyme with essential antioxidant functions in biology. Multiple factors including nitric oxide (NO) have been shown to attenuate its activity. However, the possible impact of NO in relation to the maturation of active catalase, including its heme acquisition and tetramer formation, has not been investigated. We found that NO attenuates heme insertion into catalase in both short-term and long-term incubations. The NO inhibition in catalase heme incorporation was associated with defective oligomerization of catalase, such that inactive catalase monomers and dimers accumulated in place of the mature tetrameric enzyme. We also found that GAPDH plays a key role in mediating these NO effects on the structure and activity of catalase. Moreover, the NO sensitivity of catalase maturation could be altered up or down by manipulating the cellular expression level or activity of thioredoxin-1, a known protein-SNO denitrosylase enzyme. In a mouse model of allergic inflammatory asthma, we found that lungs from allergen-challenged mice contained a greater percentage of dimeric catalase relative to tetrameric catalase in the unchallenged control, suggesting that the mechanisms described here are in play in the allergic asthma model. Together, our study shows how maturation of active catalase can be influenced by NO, S-nitrosylated GAPDH, and thioredoxin-1, and how maturation may become compromised in inflammatory conditions such as asthma.
authors: Department of Pathobiology, Lerner Research Institute, 9500 Euclid Avenue, Cleveland Clinic, Cleveland, OH 44195, USA.
authors: Ritu
authors: R
authors: Chakravarti
authors: School of Medicine, Case Western Reserve University, Cleveland, OH 44195, USA.
authors: Karishma
authors: K
authors: Gupta
authors: Department of Pathobiology, Lerner Research Institute, 9500 Euclid Avenue, Cleveland Clinic, Cleveland, OH 44195, USA.
authors: Alana
authors: A
authors: Majors
authors: Department of Pathobiology, Lerner Research Institute, 9500 Euclid Avenue, Cleveland Clinic, Cleveland, OH 44195, USA.
authors: Lisa
authors: L
authors: Ruple
authors: Department of Pathobiology, Lerner Research Institute, 9500 Euclid Avenue, Cleveland Clinic, Cleveland, OH 44195, USA.
authors: Mark
authors: M
authors: Aronica
authors: Department of Pathobiology, Lerner Research Institute, 9500 Euclid Avenue, Cleveland Clinic, Cleveland, OH 44195, USA. Electronic address: stuehrd@ccf.org.
authors: Dennis J
authors: DJ
authors: Stuehr
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D001249:Asthma / Q000473:pathology; D002374:Catalase / Q000378:metabolism*; D002460:Cell Line; D004195:Disease Models, Animal; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000378:metabolism*; D057809:HEK293 Cells; D006418:Heme / Q000378:metabolism*; D006801:Humans; D008168:Lung / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D032389:Myocytes, Smooth Muscle / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D010047:Ovalbumin; D013879:Thioredoxins / Q000096:biosynthesis / Q000378:metabolism*
chemical_list: D000975:Antioxidants; D009569:Nitric Oxide; D006418:Heme; D013879:Thioredoxins; D010047:Ovalbumin; D002374:Catalase; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)
keywords: Asthma; Catalase; Heme; Nitric oxide; Oligomerization; S-nitrosylated GAPDH; Thioredoxin-1