The central Z-disk region of titin is assembled from a novel repeat in variable copy numbers.
The giant sarcomeric protein titin (also described as connectin) is composed mainly of immunoglobulin (Ig)-like and fibronectin type III (fn3)-like domains arranged consecutively. At both ends of the molecule, these domains are interrupted by sequence insertions. The amino terminus of titin is localized in the Z-disk, a structure of great variability in different muscle types. We have determined the ultrastructural position of sequences in this region of the molecule in skeletal and cardiac muscle by immunoelectron microscopy using antibodies directed against unique epitopes. Titin molecules entering the Z-disk from two half sarcomeres do not significantly overlap, showing that the amino terminus is at the centre of the Z-disk. A serine/proline rich site, which can be phosphorylated by kinases in developing muscle tissues, was identified near the amino terminus of titin. Sequence analysis revealed the presence of a novel 45 residue repeat ('Z-repeats') in this region of the molecule. The number of titin Z-repeats varies due to differential splicing. We propose that this mechanism is a means of assembling Z-disks of variable thickness and mechanical strength.
Gautel
M
M
European Molecular Biology Laboratory, Structural Biology Division, Heidelberg, Germany.
Goulding
D
D
Bullard
B
B
Weber
K
K
Fürst
D O
DO
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D064211:Connectin; D018076:DNA, Complementary; D018604:Epitope Mapping; D018628:Gene Dosage; D014644:Genetic Variation; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000737:chemistry / Q000235:genetics*; D011392:Proline; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics*; D011817:Rabbits; D012091:Repetitive Sequences, Nucleic Acid; D017386:Sequence Homology, Amino Acid; D012694:Serine
D064211:Connectin; D018076:DNA, Complementary; D009124:Muscle Proteins; C577285:TTN protein, human; D012694:Serine; D011392:Proline; D011494:Protein Kinases