1H and 15N NMR resonance assignments and secondary structure of titin type I domains.
Titin/connectin is a giant muscle protein with a highly modular architecture consisting of multiple repeats of two sequence motifs, named type I and type II. Type I modules have been suggested to be intracellular members of the fibronectin type III (Fn3) domain family. Along the titin sequence they are exclusively present in the region of the molecule located in the sarcomere A-band. This region has been shown to interact with myosin and C-protein. One of the most noticeable features of type I modules is that they are particularly rich in semiconserved prolines, since these residues account for about 8% of their sequence. We have determined the secondary structure of a representative type I domain (A71) by 15N and 1H NMR. We show that the type I domains of titin have the Fn3 fold as proposed, consisting of a three- and a four-stranded beta-sheet. When the two sheets are placed on top of each other to form the beta-sandwich characteristic of the Fn3 fold, 8 out of 10 prolines are found on the same side of the molecule and form an exposed hydrophobic patch. This suggests that the semiconserved prolines might be relevant for the function of type I modules, providing a surface for binding to other A-band proteins. The secondary structure of A71 was structurally aligned to other extracellular Fn3 modules of known 3D structure. The alignment shows that titin type I modules have closest similarity to the first Fn3 domain of Drosophila neuroglian.
Muhle-Goll
C
C
European Molecular Biology Laboratory, Heidelberg, Germany.
Nilges
M
M
Pastore
A
A
D000595:Amino Acid Sequence; D000818:Animals; D064211:Connectin; D004330:Drosophila; D006859:Hydrogen; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000737:chemistry*; D009584:Nitrogen; D011494:Protein Kinases / Q000737:chemistry*; D017433:Protein Structure, Secondary; D016415:Sequence Alignment
D064211:Connectin; D009124:Muscle Proteins; D006859:Hydrogen; D011494:Protein Kinases; D009584:Nitrogen