ABSTRACT
The structure of neuromedin C, a 10-residue bombesin-like neuropeptide with the sequence Gly-Asn-His-Trp-Ala-Val-Gly-His-Leu-Met-NH2, has been investigated. Like human serum albumin, neuromedin C contains the amino-terminal CuII-, NiII-binding (ATCUN) motif which has high affinity for CuII and NiII. The solution structure of the NiII-peptide complex has been calculated based on 2D ROESY data obtained at 25 degrees C, using a hybrid distance geometry-simulated annealing approach. Comparison of 1H, 13C and 15N chemical shifts and ROESY data in the presence and absence of NiII demonstrates that the metal binds at the N-terminus of the peptide, leading to a conformational change. The metal complex adopts a conformation comprising two connected turns including residues 1Gly to 3His and 5Ala to 8His. The first turn corresponds to the NiII coordination ligands in a square planar conformation, and the second reflects the interaction between 4Trp and 8His. The results may have important physiological implications in the phenomenon of neurotransmission.
Subject(s)
Bombesin/chemistry , Copper/metabolism , Nickel/metabolism , Peptide Fragments/chemistry , Amino Acid Sequence , Bombesin/metabolism , Magnetic Resonance Spectroscopy , Neurotransmitter Agents/chemistry , Neurotransmitter Agents/metabolism , Peptide Fragments/metabolism , Protein ConformationABSTRACT
Phosphorus-31 NMR has been applied to the characterization of terminal phosphates on fragments of calf thymus DNA induced by three different nuclease systems: DNase I, DNase II and the artificial nuclease 'Mn-TMPyP/KHSO5'. In this last case, the oxidative damage to deoxyribose leads to two monophosphates esters (at the 3' and 5' ends) on both sides of the cleavage site. This method constitutes a promising approach to visualise the phosphate termini generated in DNA or RNA cleavage by cytotoxic drugs or chemical nucleases and provides a novel insight into the molecular aspects of their mechanism of action.