Open AccessImmunological detection of translation products of type V/XI collagen α 1 gene and their degradation products in red sea bream muscle
Author(s) -
TOUHATA KEN,
TOKUDA YUKI,
SAKAGUCHI MORIHIKO,
TOYOHARA HARUHIKO
Publication year - 2002
Publication title -
fisheries science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.412
H-Index - 64
eISSN - 1444-2906
pISSN - 0919-9268
DOI - 10.1046/j.1444-2906.2002.00541.x
Subject(s) - complementary dna , microbiology and biotechnology , antiserum , western blot , antibody , biochemistry , dot blot , molecular mass , type i collagen , biology , recombinant dna , chemistry , gene , enzyme , endocrinology , immunology
: We previously cloned cDNA of type V/XI collagen α1 chain (ColVa1) gene from cultured cells derived from red sea bream embryo. We raised an antibody against the deduced C‐telopeptide of ColVa1 in order to detect the translation products of this cDNA and their degradation products in red sea bream muscle. To improve its specificity, the antibody was purified from rabbit antiserum by use of an affinity column cross‐linked with recombinant C‐terminal peptide of ColVa1 produced by Epicurian coli. The purified antibody recognized a band corresponding to the α chain of type V/XI collagen in western blot analysis of the extract of cultured cells. The antibody also recognized two bands in acid‐soluble and pepsin‐solubilized collagens, indicating that the translation products of the ColVa1 gene are present in muscle and that bands correspond to α and β chains of type V/XI collagen. A band corresponding to a molecular weight of approximately 65 k was detected in the NaOH extracts of muscle, suggesting that type V/XI collagen α1 chain is restrictedly digested in red sea bream muscle.