Secondary structure determination by NMR spectroscopy of an immunoglobulin-like domain from the giant muscle protein titin.
We present the complete 15N and 1H NMR assignment and the secondary structure of an immunoglobulin-like domain from the giant muscle protein titin. The assignment was obtained using homonuclear and 15N heteronuclear 2D and 3D experiments. The complementarity of 3D TOCSY-NOESY and 3D 15N NOESY-HSQC experiments, using WATERGATE for water suppression, allowed an efficient assignment of otherwise ambiguous cross peaks and was helpful in overcoming poor TOCSY transfer for some amino acids. The secondary structure is derived from specific NOEs between backbone alpha- and amide protons, secondary chemical shifts of alpha-protons and chemical exchange for the backbone amide protons. It consists of eight beta-strands, forming two beta-sheets with four strands each, similar to the classical beta-sandwich of the immunoglobulin superfamily, as previously predicted by sequence analysis. Two of the beta-strands are connected by type II beta-turns; the first beta-strand forms a beta-bulge. The whole topology is very similar to the only intracellular immunoglobulin-like domain for which a structure has been determined so far, i.e., telokin.
Pfuhl
M
M
European Molecular Biology Laboratory, Heidelberg, Germany.
Gautel
M
M
Politou
A S
AS
Joseph
C
C
Pastore
A
A
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D064211:Connectin; D006801:Humans; D007136:Immunoglobulins / Q000737:chemistry; D007536:Isomerism; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009124:Muscle Proteins / Q000737:chemistry* / Q000235:genetics; D009219:Myosin-Light-Chain Kinase; D010446:Peptide Fragments; D010455:Peptides; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics; D017433:Protein Structure, Secondary
D000596:Amino Acids; D064211:Connectin; D007136:Immunoglobulins; D009124:Muscle Proteins; D010446:Peptide Fragments; D010455:Peptides; C577285:TTN protein, human; C060640:telokin; D011494:Protein Kinases; D009219:Myosin-Light-Chain Kinase