De Backer Joey, Maric Darko, Bosman Matthias, Dewilde Sylvia, Hoogewijs David (2021), A reliable set of reference genes to normalize oxygen-dependent cytoglobin gene expression levels in melanoma, in
Scientific Reports, 11(1), 10879-10879.
Koay Teng Wei, Osterhof Carina, Orlando Ilaria M.C., Keppner Anna, Andre Daniel, Yousefian Schayan, Alonso María Suárez, Correia Miguel, Markworth Robert, Schödel Johannes, Hankeln Thomas, Hoogewijs David (2021), Androglobin gene expression patterns and FOXJ1-dependent regulation indicate its functional association with ciliogenesis, in
Journal of Biological Chemistry, 100291-100291.
Hammerschmid Dietmar, Germani Francesca, Drusin Salvador I., Fagnen Charline, Schuster Claudio D., Hoogewijs David, Marti Marcelo A., Venien-Bryan Catherine, Moens Luc, Van Doorslaer Sabine, Sobott Frank, Dewilde Sylvia (2021), Structural modeling of a novel membrane-bound globin-coupled sensor in Geobacter sulfurreducens, in
Computational and Structural Biotechnology Journal, 19, 1874-1888.
Keppner Anna, Maric Darko, Correia Miguel, Koay Teng Wei, Orlando Ilaria M.C., Vinogradov Serge N., Hoogewijs David (2020), Lessons from the post-genomic era: Globin diversity beyond oxygen binding and transport, in
Redox Biology, 37, 101687-101687.
Randi Elisa B., Vervaet Benjamin, Tsachaki Maria, Porto Elena, Vermeylen Stijn, Lindenmeyer Maja T., Thuy Le Thi Thanh, Cohen Clemens D., Devuyst Olivier, Kistler Andreas D., Szabo Csaba, Kawada Norifumi, Hankeln Thomas, Odermatt Alex, Dewilde Sylvia, Wenger Roland H., Hoogewijs David (2020), The Antioxidative Role of Cytoglobin in Podocytes: Implications for a Role in Chronic Kidney Disease, in
Antioxidants & Redox Signaling, 32(16), 1155-1171.
Keppner Anna, Maric Darko, Sergi Chloé, Ansermet Camille, De Bellis Damien, Kratschmar Denise V., Canonica Jérémie, Klusonova Petra, Fenton Robert A., Odermatt Alex, Crambert Gilles, Hoogewijs David, Hummler Edith (2019), Deletion of the serine protease CAP2/Tmprss4 leads to dysregulated renal water handling upon dietary potassium depletion, in
Scientific Reports, 9(1), 19540-19540.
Bracke An, Peeters Kris, Punjabi Usha, Hoogewijs David, Dewilde Sylvia (2018), A search for molecular mechanisms underlying male idiopathic infertility, in
Reproductive BioMedicine Online, 36(3), 327-339.
Bracke An, Hoogewijs David, Dewilde Sylvia (2018), Exploring three different expression systems for recombinant expression of globins: Escherichia coli, Pichia pastoris and Spodoptera frugiperda, in
Analytical Biochemistry, 543, 62-70.
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Background: Hemoglobin and myoglobin represent the two most-established and well-studied heme-containing oxygen binding proteins. The post-genomic era revealed the presence of additional mammalian globin types, including neuroglobin and cytoglobin. Recently the applicant and collaborating labs identified a new metazoan globin lineage, consisting of large chimeric proteins with an N-terminal cysteine protease domain and a central globin domain, named androglobins (Adgbs), because of their specific expression in testis tissue (Hoogewijs et al. 2012a). Intriguingly, this new member of the globin family is evolutionary ancient and extremely conserved. We showed that Adgb expression is associated with postmeiotic stages of spermatogenesis. Male infertility of an Adgb-deficient mouse model and higher Adgb expression levels in spermatozoa from fertile vs. infertile males underscore the functional importance of Adgb during spermatogenesis. The present project will analyse the physiological role of Adgb using a variety of in vitro and in vivo approaches with focus on spermatogenesis.Aims: The main goal of the present proposal is to explore the functional role of Adgb. To achieve this goal, in vitro experiments were designed to determine the biochemical properties of Adgb and to characterize the mechanism of Adgb expression regulation. In vivo experiments include the completion of the ongoing generation of a novel conditional Adgb knock-out mouse model and aim to elucidate the role of Adgb during spermatogenesis. Research plan: In order to gain more insights into the function of Adgb we will perform in vitro as well as in vivo experiments. We will use cell culture studies to modify expression levels of Adgb and look at potential downstream targets by applying transcriptome profiling. Functional studies will evaluate if Adgb has a potential protection function against reactive oxygen species (ROS) in these cell models. Additionally, we will explore the transcriptional regulation of Adgb using reporter gene assays and DNA methylation status-modifying agents. Physico-chemical characterization of the recombinantly expressed Adgb globin domain (wild-type and mutants) will be performed to determine the kinetic properties of ligand binding. Functionality of the protease domain will be assessed on the recombinantly expressed calpain domain (wild-type and mutants) and full-length protein. To complement the in vitro studies we will characterize a novel conditional Adgb knock-out mouse model and initially focus on the testis-specific role of Adgb, employing testes and sperm histology, sperm functionality assays, transcriptomics and interactome studies.Significance: The elucidation of the testis-specific function of Adgb is mainly a basic science research project. However, the finding of substantially decreased Adgb expression levels in spermatozoa from infertile males and the infertility of androglobin-deficient mice clearly indicates that it could have important biomedical implications with regard to the increased occurrence of male infertility in our society. Male infertility is affected by oxygenation and ROS, among many other factors, and Adgb might be involved in sensing oxygen levels or detoxification of harmful ROS. Thereby our studies will contribute to an improved understanding of male infertility.