Biophysical Reviews

, Volume 10, Issue 2, pp 191–202 | Cite as

Molluscan hemocyanin: structure, evolution, and physiology

Review

Abstract

Most molluscs have blue blood because their respiratory molecule is hemocyanin, a type-3 copper-binding protein that turns blue upon oxygen binding. Molluscan hemocyanins are huge cylindrical multimeric glycoproteins that are found freely dissolved in the hemolymph. With molecular masses ranging from 3.3 to 13.5 MDa, molluscan hemocyanins are among the largest known proteins. They form decamers or multi-decamers of 330- to 550-kDa subunits comprising more than seven paralogous functional units. Based on the organization of functional domains, they assemble to form decamers, di-decamers, and tri-decamers. Their structure has been investigated using a combination of single particle electron cryo-microsopy of the entire structure and high-resolution X-ray crystallography of the functional unit, although, the one exception is squid hemocyanin for which a crystal structure analysis of the entire molecule has been carried out. In this review, we explain the molecular characteristics of molluscan hemocyanin mainly from the structural viewpoint, in which the structure of the functional unit, architecture of the huge cylindrical multimer, relationship between the composition of the functional unit and entire tertiary structure, and possible functions of the carbohydrates are introduced. We also discuss the evolutionary implications and physiological significance of molluscan hemocyanin.

Keywords

Molluscan hemocyanin Oxygen transporter Structure Electron cryo-microscopy X-ray crystallography Glycoprotein Evolution 

Notes

Acknowledgements

The authors thank JSPS KAKENHI (26291008 and 25450298) and Regional Innovation Strategy Support Program from the Ministry of Education, Culture, Sports, Science and Technology, Japan, for the financial support. C.G. thanks the Max Planck Society for the support.

Compliance with ethical standards

Conflict of interest

Sanae Kato declares that she has no conflicts of interest. Takashi Matsui declares that he has no conflicts of interest. Christos Gatsogiannis declares that he has no conflicts of interest. Yoshikazu Tanaka declares that he has no conflicts of interest.

Ethical approval

This article does not contain any studies with human participants by any of the authors. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Financial information

SK was a recipient of the JSPS KAKENHI (25450298) and Regional Innovation Strategy Support Program from the Ministry of Education, Culture, Sports, Science and Technology, Japan. YT received support from JSPS KAKENHI (24000011, and 15KK0248) and JST, PRESTO (JPMJPR1517). TM received support from JSPS KAKENHI (16K18501).

References

  1. Altenhein B, Markl J, Lieb B (2002) Gene structure and hemocyanin isoform HtH2 from the mollusc Haliotis Tuberculata indicate early and late intron hot spots. Gene 301:53–60CrossRefPubMedGoogle Scholar
  2. Becker MI, Arancibia S, Salazar F, Campo MD, Ioannes AD. (2014) Mollusk hemocyanins as natural immunostimulants in biomedical applications. In: Duc GHT (ed) Immune response activation. InTech, Rijeka, Croatia, pp 45–72Google Scholar
  3. Bergmann S, Lieb B, Ruth P, Markl J (2006) The hemocyanin from a living fossil, the cephalopod Nautilus pompilius: protein structure, gene organization, and evolution. J Mol Evol 62:362–374CrossRefPubMedGoogle Scholar
  4. Boisset N, Mouche F (2000) Sepia officinalis hemocyanin: a refined 3D structure from field emission gun cryoelectron microscopy. J Mol Biol 296:459–472CrossRefPubMedGoogle Scholar
  5. Cuff ME, Miller KI, van Holde KE, Hendrickson WA (1998) Crystal structure of a functional unit from octopus hemocyanin. J Mol Biol 278:855–870CrossRefPubMedGoogle Scholar
  6. Fredericq L (1878) La Physiologic du poulpe commun (Octopus vulgaris). Arch Zool Exp Gén 7:535–583Google Scholar
  7. Gai Z, Matsuno A, Kato K, Kato S, Khan MRI, Shimizu T, Yoshioka T, Kato Y, Kishimura H, Kannno G, Miyabe Y, Terada T, Tanaka Y, Yao M (2015) Crystal structure of the 3.8 MDa respiratory supermolecule hemocyanin at 3.0 Å resolution. Structure 23:2204–2212CrossRefPubMedGoogle Scholar
  8. Gatsogiannis C, Hofnagel O, Markl J, Raunser S (2015) Structure of mega-hemocyanin reveals protein origami in snails. Structure 23:93–103CrossRefPubMedGoogle Scholar
  9. Gatsogiannis C, Markl J (2009) Keyhole limpet hemocyanin: 9-a CryoEM structure and molecular model of the KLH1 didecamer reveal the interfaces and intricate topology of the 160 functional units. J Mol Biol 385:963–983CrossRefPubMedGoogle Scholar
  10. Gatsogiannis C, Moeller A, Depoix F, Meissner U, Markl J (2007) Nautilus Pompilius hemocyanin: 9 a cryo-EM structure and molecular model reveal the subunit pathway and the interfaces between the 70 functional units. J Mol Biol 374:465–486CrossRefPubMedGoogle Scholar
  11. Geyer H, Wuhrer M, Resemann A, Geyer R (2005) Identification and characterization of keyhole limpet hemocyanin N-glycans mediating cross-reactivity with Schistosoma mansoni. J Biol Chem 280:40731–40748CrossRefPubMedGoogle Scholar
  12. Ghiretti-Magaldi A, Ghiretti F (1992) The pre-history of hemocyanin. The discovery of copper in the blood of molluscs. Experientia 48:971–972CrossRefGoogle Scholar
  13. Harris JR, Markl J (1999) Keyhole limpet hemocyanin (KLH): a biomedical review. Micron 30:597–623CrossRefPubMedGoogle Scholar
  14. Harris JR, Meissner U, Gebauer W, Markl J (2004) 3D reconstruction of the hemocyanin subunit dimer from the chiton Acanthochiton fascicularis. Micron 35:23–26CrossRefPubMedGoogle Scholar
  15. Idakieva K, Stoeva S, Voelter W, Cielens C (2004) Glycosylation of Rapana thomasiana hemocyanin. Comparison with other prosobranch (gastropod) hemocyanins. Comp Biochem Physiol B 138:221–228CrossRefPubMedGoogle Scholar
  16. Jaenicke E, Buchler K, Decker H, Markl J, Schroder GF (2011) The refined structure of functional unit h of keyhole limpet hemocyanin (KLH1-h) reveals disulfide bridges. IUBMB Life 63:183–187CrossRefPubMedGoogle Scholar
  17. Jaenicke E, Buchler K, Markl J, Decker H, Barends TR (2010) Cupredoxin-like domains in haemocyanins. Biochem J 426:373–378CrossRefPubMedGoogle Scholar
  18. Jaenicke E, Fraune S, May S, Irmak P, Augustin R, Meesters C, Decker H, Zimmer M (2009) Is activated hemocyanin instead of phenoloxidase involved in immune response in woodlice? Dev Comp Immunol 33(10):1055–1063CrossRefPubMedGoogle Scholar
  19. Kröger B, Vinther J, Fuches D (2011) Cephalopod origin and evolution: a congruent picture emerging from fossils, development and molecules. BioEssays 33:602–613CrossRefPubMedGoogle Scholar
  20. Lambert O, Boisset N, Taveau JC, Lamy JN (1994) Three-dimensional reconstruction from a frozen-hydrated specimen of the chiton Lepidochiton sp. hemocyanin. J Mol Biol 244:640–647CrossRefPubMedGoogle Scholar
  21. Lamy J, You V, Taveau JC, Boisset N, Lamy JN (1998) Intramolecular localization of the functional units of sepia officinalis hemocyanin by immunoelectron microscopy. J Mol Biol 284:1051–1074CrossRefPubMedGoogle Scholar
  22. Lieb B, Altenhein B, Markl J (2000) The sequence of a gastropod hemocyanin (HtH1 from Haliotis tuberculata). J Biol Chem 275:5675–5681CrossRefPubMedGoogle Scholar
  23. Lieb B, Gebauer W, Gatsogiannis C, Depoix F, Hellmann N, Harasewych MG, Strong EE, Markl J (2010) Molluscan mega-hemocyanin: an ancient oxygen carrier tuned by a ~550 kDa polypeptide. Front Zool 7:14CrossRefPubMedPubMedCentralGoogle Scholar
  24. Lieb B, Markl J (2004) Evolution of molluscan hemocyanins as deduced from DNA sequencing. Micron 35:117–119CrossRefPubMedGoogle Scholar
  25. Markl J (2013) Evolution of molluscan hemocyanin structures. Biochim Biophys Acta 1834:1840–1852CrossRefPubMedGoogle Scholar
  26. Matsuno A, Gai Z, Tanaka M, Kato K, Kato S, Katoh T, Shimizu T, Yoshioka T, Kishimura H, Tanaka Y et al (2015) Crystallization and preliminary X-ray crystallographic study of a 3.8-MDa respiratory supermolecule hemocyanin. J Struct Biol 190:379–382CrossRefPubMedGoogle Scholar
  27. Meissner U, Dube P, Harris JR, Stark H, Markl J (2000) Structure of a molluscan hemocyanin didecamer (HtH1 from Haliotis tuberculata) at 12 a resolution by cryoelectron microscopy. J Mol Biol 298:21–34CrossRefPubMedGoogle Scholar
  28. Meissner U, Gatsogiannis C, Moeller A, Depoix F, Harris JR, Markl J (2007) Comparative 11 Å structure of two molluscan hemocyanins from 3D cryo-electron microscopy. Micron 38:754–765CrossRefPubMedGoogle Scholar
  29. Miller KI, Cuff ME, Lang WF, Varga-Weisz P, Field KG, van Holde KE (1998) Sequence of the Octopus dofleini hemocyanin subunit: structural and evolutionary implications. J Mol Biol 278:827–842CrossRefPubMedGoogle Scholar
  30. Miller KI, Schabtach E, van Holde KE (1990) Arrangement of subunits and domains within the Octopus dofleini hemocyanin molecule. Proc Natl Acad Sci USA 87:1496–1500CrossRefPubMedPubMedCentralGoogle Scholar
  31. Perbandt M, Guthohrlein EW, Rypniewski W, Idakieva K, Stoeva S, Voelter W, Genov N, Betzel C (2003) The structure of a functional unit from the wall of a gastropod hemocyanin offers a possible mechanism for cooperativity. Biochemistry 42:6341–6346CrossRefPubMedGoogle Scholar
  32. Salvini-Plawen LV, Steiner G (1996) Synapomorphies and plesiomorphies in higher classification of Mollusca. In: Taylor JD (ed) Origin and evolutionary radiation of the Mollusca. Oxford University Press, Oxford, pp 29–51Google Scholar
  33. Scheltema AH (1996) Phylogenetic position of Sipuncula, Mollusca and the progenetic Aplacophora. In: Taylor JD (ed) Origin and evolutionary radiation of the Mollusca. Oxford University Press, Oxford, pp 53–58Google Scholar
  34. Smith SA, Wilson NG, Goetz FE, Feehery C, Andrade SCS, Rouse GW, Giribet G, Dunn CW (2011) Resolving the evolutionary relationships of molluscs with phylogenomic tools. Nature 480(7377):364–367Google Scholar
  35. Siddiqui NI, Idakieva K, Demarsin B, Doumanova L, Compernolle F, Gielens C (2007) Involvement of glycan chains in the antigenicity of Rapana thomasiana hemocyanin. Biochem Biophys Res Commun 361:705–711CrossRefPubMedGoogle Scholar
  36. Siddiqui NI, Yigzaw Y, Préaux G, Gielens C (2009) Involvement of glycans in the immunological cross-reaction between α-macroglobulin and hemocyanin of the gastropod Helix pomatia. Biochimie 91:508–516CrossRefPubMedGoogle Scholar
  37. Stoeva S, Idakieva K, Betzel C, Genov N, Voelter W (2002) Amino acid sequence and glycosylation of functional unit RtH2-e from Rapana thomasiana (gastropod) hemocyanin. Arch Biochem Biophys 399:149–158CrossRefPubMedGoogle Scholar
  38. Stoeva S, Schutz J, Gebauer W, Hundsdorfer T, Manz C, Markl J, Voelter W (1999) Primary structure and unusual carbohydrate moiety of functional unit 2-c of keyhole limpet hemocyanin (KLH). Biochim Biophys Acta 1435:94–109CrossRefPubMedGoogle Scholar
  39. Thonig A, Oellermann M, Lieb B, Mark FC (2014) A new haemocyanin in cuttlefish (Sepia officinalis) eggs: sequence analysis and relevance during ontogeny. EvoDevo 5:6CrossRefPubMedPubMedCentralGoogle Scholar
  40. Waller TR (1998) Origin of the molluscan class Bivalvia and a phylogeny of major groups. In: Johnston PA, Haggart JW (eds) Bivalves: an eon of evolution. Univ. Calgary Press, Calgary, pp 1–45Google Scholar
  41. Yoshioka T, Kato S, Okamoto A (2012) Short-term rearing conditions for shipping live squids. In: Fukuta Y, Watabe S (eds) Quality improvement of coastal fish and marine invertebrates—achievement by short-term rearing and associated systems for transportation and marketing. Kouseisyakouseikaku, Tokyo, pp 106–129Google Scholar
  42. Zhang Q, Dai X, Cong Y, Zhang J, Chen DH, Dougherty MT, Wang J, Ludtke SJ, Schmid MF, Chiu W (2013) Cryo-EM structure of a molluscan hemocyanin suggests its allosteric mechanism. Structure 21:604–613CrossRefPubMedPubMedCentralGoogle Scholar
  43. Zhu H, Zhuang J, Feng H, Liang R, Wang J, Xie L, Zhu P (2014) Cryo-EM structure of isomeric molluscan hemocyanin triggered by viral infection. PLoS One 9:e98766CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© International Union for Pure and Applied Biophysics (IUPAB) and Springer-Verlag GmbH Germany, part of Springer Nature 2017

Authors and Affiliations

  1. 1.Faculty of FisheriesKagoshima UniversityKagoshimaJapan
  2. 2.Graduate School of Life SciencesTohoku UniversitySendaiJapan
  3. 3.Department of Structural BiochemistryMax Planck Institute Molecular PhysiologyDortmundGermany
  4. 4.Precursory Research for Embryonic Science and Technology (PRESTO)Japan Science and Technology Agency (JST)SendaiJapan

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