title: A 68 residue N-terminal fragment of pro-atrial natriuretic peptide is a monomeric intrinsically unstructured protein.
abstract: The mature pro forms of the cardiac natriuretic peptides, atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP), are proteolytically processed to their active hormone forms (28 and 32 residues, respectively) and N-terminal (NT)-pro fragments (68 and 76 residues, respectively). Far-ultraviolet circular dichroism (UV CD), 1D and 2D-homonuclear nuclear magnetic resonance (NMR), size exclusion-high performance liquid chromatography (SE-HPLC) and analytical ultracentrifuge sedimentation equilibrium (AUCSE) data are obtained for NT-proANP. CD data showed a large negative molar ellipticity for NT-proANP of -14,800° cm(2)/dmol at 199-200 nm. The intensity of the 1D-(1)H NMR spectra in the amide region for NT-proANP was very low and confined to ~8-8.6 ppm. Furthermore, cross-correlation resonance peaks were absent in the corresponding 2D-(1)H NOE spectra for NT-proANP in this region. The elution peak for this fragment from a G2000SW size-exclusion column was 20.4'; myoglobin (~17 K) was also eluted at 20.4'. No higher molecular weight oligomers were evident in the AUCSE experiments for NT-proANP. Collectively, the physical data demonstrate that NT-proANP, like NT-proBNP, is primarily a disordered, monomeric protein. Lastly, we compare the predictions from two in silico metaserver disorder algorithms, MeDor and MetaPrDOS, to the experimental data.
authors.affiliation: Department of Pathology and Immunology, Division of Laboratory and Genomic Medicine, Washington University School of Medicine, St Louis, MO 63110, USA. crimmins@pathology.wustl.edu
authors.forename: Dan L
authors.initials: DL
authors.lastname: Crimmins
authors.forename: Jeffrey L-F
authors.initials: JL
authors.lastname: Kao
mesh_terms: D000595:Amino Acid Sequence; D009320:Atrial Natriuretic Factor / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002942:Circular Dichroism / Q000379:methods; D003198:Computer Simulation; D006801:Humans; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000737:chemistry*; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary
chemical_list: C099254:N-terminal proatrial natriuretic peptide; D011498:Protein Precursors; D009320:Atrial Natriuretic Factor