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Here, we show that other lungfish species (<jats:italic>Protopterus\u00a0dolloi<\/jats:italic>,<jats:italic> Protopterus\u00a0aethiopicus<\/jats:italic> and <jats:italic>Lepidosiren\u00a0paradoxa<\/jats:italic>) display a similar diversity of Mb genes and have orthologous Mb genes. To investigate the functional diversification of these genes, we studied the structures, O<jats:sub>2<\/jats:sub> binding properties and nitrite reductase enzymatic activities of recombinantly expressed <jats:italic>P.\u00a0annectens<\/jats:italic> Mbs (PanMbs). CD spectroscopy and small\u2010angle X\u2010ray scattering revealed the typical globin\u2010fold in all investigated recombinant Mbs, indicating a conserved structure. The highest O<jats:sub>2<\/jats:sub> affinity was measured for PanMb2 (P<jats:sub>50<\/jats:sub>\u00a0=\u00a00.88\u00a0Torr at 20\u00a0\u00b0C), which is mainly expressed in the brain, whereas the muscle\u2010specific PanMb1 has the lowest O<jats:sub>2<\/jats:sub> affinity (P<jats:sub>50<\/jats:sub>\u00a0=\u00a03.78\u00a0Torr at 20\u00a0\u00b0C), suggesting that tissue\u2010specific O<jats:sub>2<\/jats:sub> requirements have resulted in the emergence of distinct Mb types. Two of the mainly neuronally expressed Mbs (PanMb3 and PanMb4b) have the highest nitrite reductase rates. These data show different O<jats:sub>2<\/jats:sub> binding and enzymatic properties of lungfish Mbs, reflecting multiple subfunctionalisation and neofunctionalisation events that occurred early in the evolution of lungfish. Some Mbs may have also taken over the functions of neuroglobin and cytoglobin, which are widely expressed in vertebrates but appear to be missing in lungfish.<\/jats:p>","DOI":"10.1111\/febs.15094","type":"journal-article","created":{"date-parts":[[2019,10,14]],"date-time":"2019-10-14T19:09:01Z","timestamp":1571080141000},"page":"1598-1611","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Genetic and functional diversity of the multiple lungfish myoglobins"],"prefix":"10.1111","volume":"287","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3361-5373","authenticated-orcid":false,"given":"Julia","family":"L\u00fcdemann","sequence":"first","affiliation":[{"name":"Institute of Zoology Department of Biology University of Hamburg 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