title: Carbon-13 NMR studies of the lysine side chains of calmodulin and its proteolytic fragments.
abstract: The pH-titration and dynamic behaviour of the seven lysine side chains in bovine calmodulin were studied by carbon-13 NMR. The amino groups of the calcium saturated protein and its proteolytic fragments TR1C (1-75) and TR2C (78-148) were dimethylated with carbon-13 labeled formaldehyde; this modification did not alter the protein's structure or its ability to activate the enzyme cyclic nucleotide phosphodiesterase. Tentative assignments for 5 out of the 7 dimethyl lysine resonances could be obtained by comparing spectra of the fully and partially modified protein, with those of the proteolytic fragments. The pKa values measured for calcium saturated calmodulin ranged between 9.5 (Lys 75) and 10.2 (Lys 13); two residues (Lys 94 and Lys 13) showed a biphasic titration curve suggesting their possible involvement in ion-pairs. The dynamic behavior of the lysine side chains was deduced from spin lattice relaxation measurements. All side chains were flexible and this was not influenced by the removal of calcium, or the addition of the calmodulin antagonist trifluoperazine. The latter data suggest that the lysine side chains are not directly involved in calmodulin's target binding sites.
authors.affiliation: Department of Biological Sciences, University of Calgary, Canada.
authors.forename: M E
authors.initials: ME
authors.lastname: Huque
authors.forename: H J
authors.initials: HJ
authors.lastname: Vogel
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002147:Calmodulin / Q000737:chemistry* / Q000302:isolation & purification; D002247:Carbon Isotopes; D002417:Cattle; D008239:Lysine / Q000031:analogs & derivatives / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008297:Male; D008745:Methylation; D010446:Peptide Fragments / Q000737:chemistry*; D011487:Protein Conformation; D013737:Testis / Q000378:metabolism; D014357:Trypsin
chemical_list: D002147:Calmodulin; D002247:Carbon Isotopes; D010446:Peptide Fragments; D014357:Trypsin; D008239:Lysine