Open AccessA 49 kDa microtubule cross‐linking protein from Artemia franciscana is a coenzyme A‐transferase
Author(s) -
Oulton Mindy M.,
Amons Reinout,
Liang Ping,
MacRae Thomas H.
Publication year - 2003
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1033.2003.03898.x
Subject(s) - biology , edman degradation , biochemistry , peptide sequence , genetics , brine shrimp , transferase , gene , microbiology and biotechnology , enzyme , ecology
Embryos and larvae of the brine shrimp, Artemia franciscana , were shown previously to possess a protein, now termed p49, which cross‐links microtubules in vitro . Molecular characteristics of p49 were described, but the protein's identity and its role in the cell were not determined. Degenerate oligonucleotide primers designed on the basis of peptide sequence obtained by Edman degradation during this study were used to generate p49 cDNAs by RT‐PCR and these were cloned and sequenced. Comparison with archived sequences revealed that the deduced amino acid sequence of p49 resembled the Drosophila gene product CG7920, as well as related proteins encoded in the genomes of Anopheles and Caenorhabditis . Similar proteins exist in several bacteria but no evident homologues were found in vertebrates and plants, and only very distant homologues resided in yeast. When evolutionary relationships were compared, p49 and the homologues from Drosophila , Anopheles and Caenorhabditis formed a distinct subcluster within phylogenetic trees. Additionally, the predicted secondary structures of p49, 4‐hydroxybutyrate CoA‐transferase from Clostridium aminobutyricum and glutaconate CoA‐transferase from Acidaminococcus fermentans were similar and the enzymes may possess related catalytic mechanisms. The purified Artemia protein exhibited 4‐hydroxybutyrate CoA‐transferase activity, thereby establishing p49 as the first crustacean CoA‐transferase to be characterized. Probing of Western blots with an antibody against p49 revealed a cross‐reactive protein in Drosophila that associated with microtubules, but to a lesser extent than did p49 from Artemia .