title: Random coil conformation of a Gly/Ala-rich insert in IkappaB alpha excludes structural stabilization as the mechanism for protection against proteasomal degradation.
abstract: Peptide segments of multiple glycine and alanine residues prevent the proteolytic degradation of ubiquitinated proteins by the proteasome. The structure of a Gly/Ala-rich insert in IkappaB alpha was probed by nuclear magnetic resonance (NMR) spectroscopy, comparing IkappaB alpha samples with and without Gly/Ala-rich insert. Narrow 1H-NMR resonances at chemical shifts indicative of random coil conformations were observed in the difference spectrum. circular dichroism (CD) measurements further confirm that the mechanism of protection against proteolytic degradation is not based on structural transition or stabilization caused by the Gly/Ala-rich segment. In addition, most of the N- and C-terminal residues outside the ankyrin repeats in wild-type IkappaB alpha were found to be flexibly disordered.
authors.affiliation: Microbiology and Tumor Biology Center (MTC), Karolinska Institute, Stockholm, Sweden.
authors.forename: A
authors.initials: A
authors.lastname: Leonchiks
authors.forename: E
authors.initials: E
authors.lastname: Liepinsh
authors.forename: M
authors.initials: M
authors.lastname: Barishev
authors.forename: A
authors.initials: A
authors.lastname: Sharipo
authors.forename: M G
authors.initials: MG
authors.lastname: Masucci
authors.forename: G
authors.initials: G
authors.lastname: Otting
mesh_terms: D000409:Alanine / Q000737:chemistry; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D005998:Glycine / Q000737:chemistry; D020672:I-kappa B Proteins; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000378:metabolism*; D000072000:NF-KappaB Inhibitor alpha; D010455:Peptides / Q000737:chemistry; D018583:Pliability; D046988:Proteasome Endopeptidase Complex; D011487:Protein Conformation
chemical_list: D000596:Amino Acids; D004268:DNA-Binding Proteins; D020672:I-kappa B Proteins; D009097:Multienzyme Complexes; D010455:Peptides; D000072000:NF-KappaB Inhibitor alpha; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D000409:Alanine; D005998:Glycine