Molecular Biology Reports

, Volume 37, Issue 4, pp 2009–2019

Molecular cloning and localization of a calpain-like protease from the abdominal muscle of Norway lobster Nephrops norvegicus

  • S. G. Gornik
  • G. D. Westrop
  • G. H. Coombs
  • D. M. Neil
Article

DOI: 10.1007/s11033-009-9652-x

Cite this article as:
Gornik, S.G., Westrop, G.D., Coombs, G.H. et al. Mol Biol Rep (2010) 37: 2009. doi:10.1007/s11033-009-9652-x

Abstract

Calpains are ubiquitous cysteine-proteases found in many, if not all, living organisms and their roles within these organisms are diverse, ranging from the mediation of cytoskeletal remodeling to the regulation of gene expression. In crustaceans calpains have so far been shown to be important mainly during moulting and growth. In the present study we report the expression of a calpain in the abdominal muscle of Norway lobster (Nephrops norvegicus) using degenerate primer, rapid amplification of cDNA ends (5′-3′-RACE), reverse transcriptase-PCR and RNA in situ hybridization approaches. The full-length mRNA sequence (2,774 bp) was found to include an open reading frame (bp 225–1,940) encoding a 572 amino acid polypeptide with a predicted mass of 65.9 kDa and a predicted pI of 5.17. The calpain was found to be an arthropod M-class calpain homologue to Homarus americanus Calpain M (Ha-CalpM) and has thus been termed Nephrops norvegicus calpain M (Nn-CalpM). When its expression pattern in abdominal muscle of adult intermoult Nephrops norvegicus was investigated an exclusive expression in a thin layer of connective tissue cells surrounding muscle fibres was found. This localization suggests a role in tenderizing connective tissue networks during growth and moulting.

Keywords

Calpain M Ca2+-dependent cysteine protease Nephrops norvegicus Crustacean moult 

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • S. G. Gornik
    • 1
    • 3
  • G. D. Westrop
    • 2
  • G. H. Coombs
    • 2
  • D. M. Neil
    • 1
  1. 1.Division of Ecology and Evolutionary Biology (DEEB), Faculty of Biomedical and Life SciencesUniversity of GlasgowGlasgowScotland, UK
  2. 2.Strathclyde Institute of Pharmacy and Biomedical SciencesUniversity of StrathclydeGlasgowScotland, UK
  3. 3.School of BotanyUniversity of MelbourneParkvilleAustralia

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