title: Backbone dynamics of human parathyroid hormone (1-34): flexibility of the central region under different environmental conditions.
abstract: The presence of a stable tertiary structure in the bioactive N-terminal portion of parathyroid hormone (PTH), a major hormone in the maintenance of extracellular calcium homeostasis, is still debated. In this work, 15N relaxation parameters of the 33 backbone amides of human PTH(1-34) were determined in phosphate-buffered saline solution (PBS) and in the presence of dodecylphosphocholine (DPC) micelles. The relaxation parameters were analyzed using both the model-free formalism (G. Lipari and A. Szabo, Journal of the American Chemical Society, 1982, Vol. 104, pp. 4546-4549) and the reduced spectral density functions approach (J.-F. Lefevre, K. T. Dayie, J. W. Peng, and G. Wagner, Biochemistry, 1996, Vol. 35, pp. 2674-2686). In PBS, the region around Gly12 possesses a high degree of flexibility and the C-terminal helix is less flexible than the N-terminal one. In the presence of DPC micelles, the mobility of the entire molecule is reduced, but the stability of the N-terminal helix increases relative to the C-terminal one. A point of relatively higher mobility at residue Gly12 is still present and a new site of local mobility at residues 16-17 is generated. These results justify the lack of experimental nuclear Overhauser effect (NOE) restraints with lack of tertiary structure and support the hypothesis that, in the absence of the receptor, the relative spatial orientation of the two N- and C-terminal helices is undefined. The flexibility in the midregion of PTH(1-34), maintained in the presence of the membrane-mimetic environment, may enable the correct relative disposition of the two helices, favoring a productive interaction with the receptor.
authors.affiliation: Institute of Biomolecular Chemistry, CNR, Department of Chemical Sciences, University of Padova, via Marzolo 1, 35131 Padova, Italy.
authors.forename: Michele
authors.initials: M
authors.lastname: Scian
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Marin
authors.forename: Massimo
authors.initials: M
authors.lastname: Bellanda
authors.forename: Liqiang
authors.initials: L
authors.lastname: Tou
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Alexander
authors.forename: Michael
authors.initials: M
authors.lastname: Rosenblatt
authors.forename: Michael
authors.initials: M
authors.lastname: Chorev
authors.forename: Evaristo
authors.initials: E
authors.lastname: Peggion
authors.forename: Stefano
authors.initials: S
authors.lastname: Mammi
mesh_terms: D000595:Amino Acid Sequence; D002021:Buffers; D004926:Escherichia coli / Q000235:genetics; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008823:Micelles; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D010281:Parathyroid Hormone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification; D010446:Peptide Fragments / Q000737:chemistry; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000737:chemistry; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012965:Sodium Chloride / Q000737:chemistry*; D012996:Solutions
chemical_list: D002021:Buffers; D008823:Micelles; D009587:Nitrogen Isotopes; C470125:PTH protein, human; D010281:Parathyroid Hormone; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D012996:Solutions; D010767:Phosphorylcholine; D012965:Sodium Chloride; C028810:dodecylphosphocholine; D005982:Glutathione Transferase