Structure of Tribolium castaneum hexamerin solved by cryo-electron microscopy

Lucie Valentová1, Tibor Füzik1, Antonín Přidal2, Pavel Plevka1

1Structural virology, Central European Institute of Technology, Masaryk University, Kamenice 5, 625 00, Brno, Czech Republic

2Department of Zoology, Fishery, Hydrobiology, and Apidology, Faculty of Agronomy, Mendel University in Brno, Zemědělská 1, 613 00, Brno, Czech Republic

Hexamerins are the most abundant proteins in larval stages of Holometabola insects. In the pupae haemolymph hexamerins constitute up to 50 % of all proteins [1]. Hexamerins evolved from crustacean hemocyanins but lost the ability to bind copper ions and to transport oxygen [2]; instead, they serve as amino acid source during the non-feeding period, when pupae undergo the development toward the adult stage. There is also evidence that hexamerins are able to bind small organic compounds and hormones such as juvenile hormone, which is involved in cuticule formation and humoral immune defence [1]. Tribolium castaneum known as a red flour beetle is a worldwide pest which causes damages of cereals, legumes and dried stored products. It has evolved resistance against a variety of insecticides [3].

Here we present the structure of native Tribolium castaneum hexamerin solved by cryo-electron microscopy. Hexamerin was purified from T. castaneum pupae and vitrified on holey carbon coated copper grids. Data were collected on FEI Titan Krios microscope operated at 300 kV. The final data analysis led to an electrostatic potential map with the resolution of 3.9 Å. Molecular structure was built into this map. Structure of hexamerin provides a basis for further studies of Tribolium life cycle.

1.         Burmester, T. (1999) Evolution and function of the insect hexamerins. European Journal of Entomology. 96(3) 213-225.

2.         Martins, J. R., Nunes, F. M. F., Cristino, A. S., Simoes, Z. P. and Bitondi, M. M. G. (2010) The four hexamerin genes in the honey bee: structure, molecular evolution and function deduced from expression patterns in queens, workers and drones. Bmc Molecular Biology. 11 20.

3          CABI. (2007) Tribolium castaneum  (red flour beetle) datasheet. Invasive Species Compendium   CAB International Publishing, Wallingford, UK.