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

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...

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Bibliographic Details
Published in:Molecular biology reports 2010-04, Vol.37 (4), p.2009-2019
Main Authors: Gornik, S. G, Westrop, G. D, Coombs, G. H, Neil, D. M
Format: Article
Language:English
Subjects:
Abdomen
Amino Acid Sequence
Animal Anatomy
Animal Biochemistry
Animals
Base Sequence
Biomedical and Life Sciences
Calpain - chemistry
Calpain - genetics
Cloning
Cloning, Molecular
Crustaceans
Digoxigenin - metabolism
DNA, Complementary - genetics
Gene Expression Regulation
Histology
Hybridization
In Situ Hybridization
Life Sciences
Male
Molecular biology
Molecular Sequence Data
Morphology
Muscles - cytology
Muscles - enzymology
Nephropidae - enzymology
Norway
Phylogeny
Proteases
Protein Transport
Reverse Transcriptase Polymerase Chain Reaction
Sequence Alignment
Sequence Analysis, DNA
Sequence Analysis, Protein
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Summary: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.
ISSN:0301-4851
1573-4978
DOI:10.1007/s11033-009-9652-x