Purification and physical properties of nematode mini-titins and their relation to twitchin.
We have isolated mini-titin from the nematodes Ascaris lumbricoides and Caenorhabditis elegans under native conditions using a modification in the procedure to prepare this protein from insect muscle. The proteins have an apparent molecular weight of 600,000 and appear in oriented specimens as flexible thin rods with a length around 240-250 nm. The circular dichroism spectrum of the Ascaris protein is dominated by beta-structure. The proteins react with antibodies to insect mini-titin and also with antibodies raised against peptides contained in the sequence predicted for twitchin, the product of the Caenorhabditis elegans unc-22 gene. Antibodies to insect mini-titin decorate the body musculature as well as the pharynx of wild-type C. elegans in immunofluorescence microscopy. In the twitchin mutant E66 only the pharynx is decorated. We conclude that the mini-titins of invertebrate muscles defined earlier by ultrastructural criteria are very likely to be twitchins, i.e. molecules necessary for normal muscle contraction. We discuss the molecular properties of the proteins in the light of the sequence established for twitchin.
Nave
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Max Planck Institute for Biophysical Chemistry, Department of Biochemistry, Goettingen, FRG.
Fürst
D
D
Vinkemeier
U
U
Weber
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001200:Ascaris / Q000032:analysis*; D015153:Blotting, Western; D002107:Caenorhabditis / Q000032:analysis*; D029742:Caenorhabditis elegans Proteins; D002148:Calmodulin-Binding Proteins; D002942:Circular Dichroism; D064211:Connectin; D016384:Consensus Sequence; D015801:Helminth Proteins / Q000737:chemistry*; D019476:Insect Proteins; D007447:Invertebrate Hormones / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000737:chemistry* / Q000302:isolation & purification; D013329:Structure-Activity Relationship
D029742:Caenorhabditis elegans Proteins; D002148:Calmodulin-Binding Proteins; D064211:Connectin; D015801:Helminth Proteins; D019476:Insect Proteins; D007447:Invertebrate Hormones; D009124:Muscle Proteins; C489233:unc-22 protein, C elegans