Skip to main content
SpringerLink
Log in

Molluscan hemocyanin: structure, evolution, and physiology

  • Review
  • Published:
Biophysical Reviews Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

Similar content being viewed by others

References

  • 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 301:53–60

    Article  CAS  PubMed  Google Scholar 

  • 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 45–72

  • 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 62:362–374

    Article  CAS  PubMed  Google Scholar 

  • Boisset N, Mouche F (2000) Sepia officinalis hemocyanin: a refined 3D structure from field emission gun cryoelectron microscopy. J Mol Biol 296:459–472

    Article  CAS  PubMed  Google Scholar 

  • Cuff ME, Miller KI, van Holde KE, Hendrickson WA (1998) Crystal structure of a functional unit from octopus hemocyanin. J Mol Biol 278:855–870

    Article  CAS  PubMed  Google Scholar 

  • Fredericq L (1878) La Physiologic du poulpe commun (Octopus vulgaris). Arch Zool Exp Gén 7:535–583

    Google Scholar 

  • 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 23:2204–2212

    Article  CAS  PubMed  Google Scholar 

  • Gatsogiannis C, Hofnagel O, Markl J, Raunser S (2015) Structure of mega-hemocyanin reveals protein origami in snails. Structure 23:93–103

    Article  CAS  PubMed  Google Scholar 

  • 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 385:963–983

    Article  CAS  PubMed  Google Scholar 

  • 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 374:465–486

    Article  CAS  PubMed  Google Scholar 

  • 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 280:40731–40748

    Article  CAS  PubMed  Google Scholar 

  • Ghiretti-Magaldi A, Ghiretti F (1992) The pre-history of hemocyanin. The discovery of copper in the blood of molluscs. Experientia 48:971–972

    Article  CAS  Google Scholar 

  • Harris JR, Markl J (1999) Keyhole limpet hemocyanin (KLH): a biomedical review. Micron 30:597–623

    Article  CAS  PubMed  Google Scholar 

  • Harris JR, Meissner U, Gebauer W, Markl J (2004) 3D reconstruction of the hemocyanin subunit dimer from the chiton Acanthochiton fascicularis. Micron 35:23–26

    Article  CAS  PubMed  Google Scholar 

  • Idakieva K, Stoeva S, Voelter W, Cielens C (2004) Glycosylation of Rapana thomasiana hemocyanin. Comparison with other prosobranch (gastropod) hemocyanins. Comp Biochem Physiol B 138:221–228

    Article  PubMed  Google Scholar 

  • 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 63:183–187

    Article  CAS  PubMed  Google Scholar 

  • Jaenicke E, Buchler K, Markl J, Decker H, Barends TR (2010) Cupredoxin-like domains in haemocyanins. Biochem J 426:373–378

    Article  CAS  PubMed  Google Scholar 

  • 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):1055–1063

    Article  CAS  PubMed  Google Scholar 

  • Kröger B, Vinther J, Fuches D (2011) Cephalopod origin and evolution: a congruent picture emerging from fossils, development and molecules. BioEssays 33:602–613

    Article  PubMed  Google Scholar 

  • 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 244:640–647

    Article  CAS  PubMed  Google Scholar 

  • 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 284:1051–1074

    Article  CAS  PubMed  Google Scholar 

  • Lieb B, Altenhein B, Markl J (2000) The sequence of a gastropod hemocyanin (HtH1 from Haliotis tuberculata). J Biol Chem 275:5675–5681

    Article  CAS  PubMed  Google Scholar 

  • 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 7:14

    Article  PubMed  PubMed Central  Google Scholar 

  • Lieb B, Markl J (2004) Evolution of molluscan hemocyanins as deduced from DNA sequencing. Micron 35:117–119

    Article  CAS  PubMed  Google Scholar 

  • Markl J (2013) Evolution of molluscan hemocyanin structures. Biochim Biophys Acta 1834:1840–1852

    Article  CAS  PubMed  Google Scholar 

  • 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 190:379–382

    Article  CAS  PubMed  Google Scholar 

  • 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 298:21–34

    Article  CAS  PubMed  Google Scholar 

  • 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 38:754–765

    Article  CAS  PubMed  Google Scholar 

  • 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 278:827–842

    Article  CAS  PubMed  Google Scholar 

  • Miller KI, Schabtach E, van Holde KE (1990) Arrangement of subunits and domains within the Octopus dofleini hemocyanin molecule. Proc Natl Acad Sci USA 87:1496–1500

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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 42:6341–6346

    Article  CAS  PubMed  Google Scholar 

  • 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 29–51

  • 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 53–58

  • 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):364–367

  • 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 361:705–711

    Article  CAS  PubMed  Google Scholar 

  • 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 91:508–516

    Article  CAS  PubMed  Google Scholar 

  • 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 399:149–158

    Article  CAS  PubMed  Google Scholar 

  • 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 1435:94–109

    Article  CAS  PubMed  Google Scholar 

  • Thonig A, Oellermann M, Lieb B, Mark FC (2014) A new haemocyanin in cuttlefish (Sepia officinalis) eggs: sequence analysis and relevance during ontogeny. EvoDevo 5:6

    Article  PubMed  PubMed Central  Google Scholar 

  • 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 1–45

  • 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 106–129

    Google Scholar 

  • 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 21:604–613

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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:e98766

    Article  PubMed  PubMed Central  Google Scholar 

Download references

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.

Author information

Authors and Affiliations

  1. Faculty of Fisheries, Kagoshima University, 4-50-20 Shimoarata, Kagoshima, 890-0056, Japan

    Sanae Kato

  2. Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, 980-8577, Japan

    Takashi Matsui & Yoshikazu Tanaka

  3. Department of Structural Biochemistry, Max Planck Institute Molecular Physiology, 44227, Dortmund, Germany

    Christos Gatsogiannis

  4. Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), Sendai, 980-8577, Japan

    Yoshikazu Tanaka

Authors
  1. Sanae Kato
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Takashi Matsui
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Christos Gatsogiannis
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yoshikazu Tanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Sanae Kato or Yoshikazu Tanaka.

Ethics declarations

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).

Additional information

This article is part of a Special Issue on ‘Biomolecules to Bio-nanomachines - Fumio Arisaka 70th Birthday’ edited by Damien Hall, Junichi Takagi and Haruki Nakamura.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kato, S., Matsui, T., Gatsogiannis, C. et al. Molluscan hemocyanin: structure, evolution, and physiology. Biophys Rev 10, 191–202 (2018). https://doi.org/10.1007/s12551-017-0349-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12551-017-0349-4

Keywords

Search

Navigation