Heterogeneity and a coiled coil prediction of trypanosomatid-like flagellar rod proteins in Euglena.
The emergent flagellum of euglenoids and trypanosomatids contained in addition to microtubules a prominent filamentous structure--the flagellar rod (paraflagellar/paraxonemal rod). Immunoblots and immunofluorescence localization using three antibodies generated against gel-isolated proteins confirmed previous studies that the Euglena flagellar rod consisted of polypeptides migrating at 66-, 69-, and 75-kD. Immunoblotting after two dimensional gel electrophoresis identified ten or more isoforms of these polypeptides. Differences in migration in acrylamide gels under nonreducing and reducing conditions suggested that the rod proteins contain intramolecular disulfide linkages. Comparative peptide mapping showed that the 66-, 69-, and 75-kD polypeptides were distinct, but related proteins, and also identified a fourth related protein migrating at 64-kD. Using antibodies against rod proteins, two overlapping cDNAs were isolated and from their sequences the cDNAs were predicted to encode 334 amino acids of the 66-kD protein; the amino acid sequence had > 65% identity to the carboxyl-terminus of the trypanosomatid flagellar rod proteins. Secondary structural prediction suggested that flagellar rod proteins contain an extended segmented coiled coil stalk and two nonhelical heads. Coiled coil appeared to be an important structural motif in the construction of flagellar rod filaments.
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Department of Biological Sciences (m/c 066), University of Illinois at Chicago 60607-7080, USA. huan.ngo@quickmail.yale.edu
Bouck
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D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D003599:Cytoskeleton / Q000737:chemistry; D018076:DNA, Complementary; D005056:Euglena gracilis / Q000737:chemistry* / Q000235:genetics; D005407:Flagella / Q000737:chemistry*; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D007894:Leishmania mexicana / Q000737:chemistry; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D017433:Protein Structure, Secondary; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D016415:Sequence Alignment; D014345:Trypanosoma / Q000737:chemistry
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D015800:Protozoan Proteins