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Molecular cloning and 3D structure prediction of myoglobin and cytoglobin in Eurasian Tree Sparrow Passer montanus

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Abstract

In vertebrates, myoglobin (Mb) and cytoglobin (Cygb) are closest relatives in the family of globins, which are heme-containing proteins that can bind gaseous molecules. Mb acts not only as an O2 transporter but also a nitric oxide (NO) scavenger in cardiac and striated muscle. Cygb has been suggested to play important functions in lipid-based signaling processes, defense against reactive oxygen species (ROS), and nitric oxide (NO) metabolism, and it is present in a variety of cell types. However, little information about the structures and functions of Mb and Cygb is known in birds. Here, we cloned the full-length open reading frames (ORFs) of the two globins in Eurasian Tree Sparrow (Passer montanus). The Mb ORF cDNA contains 465 base pairs (bp) encoding 154 amino acids (aa), and the Cygb ORF cDNA contains 540 bp encoding 179 aa. Our results show that the amino acid sequences and three-dimensional (3D) structures of Mb and Cygb are highly conserved in vertebrate species. Interestingly, two specific substitutions were detected in Cygb compared with other vertebrates, which resulted in slight variation of the 3D conformation (e.g., distance between Tyr H16 and Lys G8, the strength of hydrogen bonds, and angles between the G–H helices). Our results may contribute to further understanding the structures, properties, and functions of Mb and Cygb as well as the potential mechanisms of oxygen utilization pathways in vertebrates.

Zusammenfassung

Molekulare klonierung und 3D-strukturvorhersage von myoglobin und cytoglobin beim feldsperling

Bei vertebraten sind myoglobin (Mb) und cytoglobin (Cygb) nahe Verwandte in der Familie der Globine, einer Stoffgruppe, die aus Häm-haltigen Proteinen besteht, die gasförmige Moleküle binden können. Mb dient nicht nur als Sauerstofftransporter, sondern auch als Stickoxid-Reiniger im Herzmuskel und gestreiften Muskeln. Cygb ist ein neu entdecktes Globin, und es wird angenommen, dass es eine wichtige Rolle spielt in lipid-basierten Signal-Prozessen, in der Verteidigung gegen reaktive Sauerstoffspezies und im Stickoxid-Stoffwechsel, der sich in verschiedenen Zelltypen zeigt. Allerdings ist wenig über Struktur und Funktion von Mb und Cygb bei Vögeln bekannt. Wir klonierten die gesamten „open reading frames“(ORFs) der zwei Globine des Feldsperling (Passer montanus). Die Mb ORF cDNA enthält 465 Basenpaare (bp), die für 154 Aminosäuren (aa) kodieren, und die Cygb ORF cDNA enthält 540 bp, die für 179 aa kodieren. Unsere Ergebnisse zeigen, dass die Aminosäuresequenzen und 3D-Strukturen von Mb und Cygb in Wirbeltierarten hoch konserviert sind. Interessanterweise fanden wir zwei spezifische Substitutionen in Cygb im Vergleich mit anderen Vertebraten, die zu einer leichten Änderung in der 3D-Struktur führten (z.B. Abstand zwischen Tyr H16 und Lys G8, die Kraft der Wasserstoffbindungen und Winkel zwischen den G-H Helices). Unsere Ergebnisse könnten einen Beitrag leisten zur weiteren Darstellung der Strukturen, Eigenschaften und Funktionen von Mb und Cygb; und zum Verständnis von möglichen Mechanismen von Sauerstoff-Pathways bei Vertebraten.

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Acknowledgments

We thank the two anonymous reviewers whose comments and suggestions helped improve and clarify the manuscript. This study was supported by the National Natural Science Foundation of China (NSFC, 31330073, 31000191), the foundation of China Scholarship Council (CSC, 201408130068), and the Natural Science Foundation of the Department of Education, Hebei Province (YQ2014024) to D.M.L., the Natural Science Foundation of Hebei Province (NSFHB, C2013205018) to Y.F.W.

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Authors and Affiliations

  1. Key Laboratory of Animal Physiology, Biochemistry and Molecular Biology of Hebei Province, College of Life Sciences, Hebei Normal University, 20th East Road of 2nd South Ring, Yuhua District, Shijiazhuang, 050024, People’s Republic of China

    Lina Wu, Yanfeng Sun, Mo Li, Yaqing Li, Yao Yao, Xuelu Liu, Yinchao Hao, Dongming Li & Yuefeng Wu

  2. Department of Biology and Chemistry, Xingtai University, Xingtai, 054001, People’s Republic of China

    Lina Wu

  3. Ocean College of Hebei Agricultural University, Qinhuangdao, 066003, People’s Republic of China

    Yanfeng Sun

  4. Shijiazhuang Academy of Agriculture and Forestry Sciences, Shijiazhuang, 050041, People’s Republic of China

    Yaqing Li

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  1. Lina Wu
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Correspondence to Dongming Li or Yuefeng Wu.

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Communicated by K. C. Klasing.

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Wu, L., Sun, Y., Li, M. et al. Molecular cloning and 3D structure prediction of myoglobin and cytoglobin in Eurasian Tree Sparrow Passer montanus . J Ornithol 157, 493–504 (2016). https://doi.org/10.1007/s10336-015-1290-6

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