Characterization of osteocalcin (BGP) and matrix Gla protein (MGP) fish specific antibodies: validation for immunodetection studies in lower vertebrates

In fish species the basic mechanisms of bone development and bone remodeling are not fully understood. The classification of bone tissue in teleosts as cellular or acellular and the presence of transitional states between bone and cartilage and the finding of different types of cartilage in teleosts...

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Bibliographic Details
Published in:Calcified tissue international 2004-04, Vol.74 (2), p.170-180
Main Authors: Simes, D C, Williamson, M K, Schaff, B J, Gavaia, P J, Ingleton, P M, Price, P A, Cancela, M L
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Antibody Specificity - immunology
Argyrosomus regius
Bone and Bones - immunology
Bone and Bones - metabolism
Calcium-Binding Proteins - immunology
Calcium-Binding Proteins - metabolism
Electrophoresis, Polyacrylamide Gel
Extracellular Matrix Proteins
Fishes - immunology
Fishes - metabolism
Galeorhinus galeus
Matrix Gla Protein
Molecular Sequence Data
Osteocalcin - immunology
Osteocalcin - metabolism
Species Specificity
Xenopus - immunology
Xenopus - metabolism
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Summary:In fish species the basic mechanisms of bone development and bone remodeling are not fully understood. The classification of bone tissue in teleosts as cellular or acellular and the presence of transitional states between bone and cartilage and the finding of different types of cartilage in teleosts not previously recognized in higher vertebrates emphasizes the need for a study on the accumulation of the Gla-containing proteins MGP and BGP at the cellular level. In the present study, polyclonal antibodies developed against BGP and MGP from A. regius (a local marine teleost fish) and against MGP from G. galeus (a Pacific Ocean shark), were tested by Western blot for their specificity against BGP and MGP from several other species of teleost fish and shark. For this purpose we extracted and purified both proteins from various marine and freshwater teleosts, identified them by N-terminal amino acid sequence analysis and confirmed the presence of gamma-carboxylation in the proteins with the use of a stain specific for Gla residues. Each antibody recognized either BGP or MGP with no cross-reaction between proteins detected. All purified fish BGPs and MGPs tested were shown to be specifically recognized, thus validating the use of these antibodies for further studies.
ISSN:0171-967X
1432-0827
DOI:10.1007/s00223-003-0079-4