title: Binding of alpha-actinin to titin: implications for Z-disk assembly.
abstract: Titin is an exceptionally large protein (M.Wt. approximately 3 MDa) that spans half the sarcomere in muscle, from the Z-disk to the M-line. In the Z-disk, it interacts with alpha-actinin homodimers that are a principal component of the Z-filaments linking actin filaments. The interaction between titin and alpha-actinin involves repeating approximately 45 amino acid sequences (Z-repeats) near the N-terminus of titin and the C-lobe of the C-terminal calmodulin-like domain of alpha-actinin. The conformation of Z-repeat 7 (ZR7) of titin when complexed with the 73-amino acid C-terminal portion of alpha-actinin (EF34) was studied by heteronuclear NMR spectroscopy using (15)N-labeling of ZR7 and found to be helical over a stretch of 18 residues. Complex formation resulted in the protection of one site of preferential cleavage of EF34 at Phe14-Leu17, as determined by limited proteolysis experiments coupled to mass spectrometry measurements. Intermolecular NOEs show Val16 of ZR7 to be positioned close in space to the backbone of EF34 around Phe14. These observations suggest that the mode of binding of ZR7 to EF34 is similar to that of troponin I to troponin C and of peptide C20W to calmodulin. These complexes would appear to represent a general alternative binding mode of calmodulin-like domains to target peptides.
authors.affiliation: Division of Molecular Structure, National Institute for Medical Research, The Ridgeway, Mill Hill, London, United Kingdom.
authors.forename: R A
authors.initials: RA
authors.lastname: Atkinson
authors.forename: C
authors.initials: C
authors.lastname: Joseph
authors.forename: F
authors.initials: F
authors.lastname: Dal Piaz
authors.forename: L
authors.initials: L
authors.lastname: Birolo
authors.forename: G
authors.initials: G
authors.lastname: Stier
authors.forename: P
authors.initials: P
authors.lastname: Pucci
authors.forename: A
authors.initials: A
authors.lastname: Pastore
mesh_terms: D000185:Actinin / Q000737:chemistry*; D000595:Amino Acid Sequence; D064211:Connectin; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000737:chemistry*; D009587:Nitrogen Isotopes; D010446:Peptide Fragments / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D011494:Protein Kinases / Q000737:chemistry*; D017433:Protein Structure, Secondary; D016415:Sequence Alignment
chemical_list: D064211:Connectin; D009124:Muscle Proteins; D009587:Nitrogen Isotopes; D010446:Peptide Fragments; D000185:Actinin; D011494:Protein Kinases