title: Structural analysis of the small chain of the 2S albumin, napin nIII, from rapeseed. Chemical and spectroscopic evidence of an intramolecular bond formation.
abstract: Napin nIII is a 2S albumin from rapeseed (Brassica napus L.) homologous to the major mustard allergen. It is composed of two different polypeptide chains linked by two disulphide bonds. The small chain has been isolated by reverse-phase HPLC after reduction of the native protein and its primary structure elucidated. This 37 residue polypeptide contains only two cysteines, at positions 10 and 23, which show a great tendency to form a non-native intramolecular disulphide bridge. The kinetic analysis of this process was performed by measuring the fluorescence emission of the single tryptophan residue of the molecule since its fluorescence intensity is about 30% decreased during disulphide formation. Small changes on the secondary structure of the polypeptide were measured by circular dichroism. The process is delayed in the presence of the reduced large chain of nIII. However, no dimer formation was detected under the conditions used, either between small chains or between the small and the large chains. Thus, the interchain disulphide formation in napin nIII should be considered as an early step during maturation of this multi-subunit seed protein.
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad Complutense, Madrid, Spain.
authors.forename: R I
authors.initials: RI
authors.lastname: Monsalve
authors.forename: M
authors.initials: M
authors.lastname: Villalba
authors.forename: C
authors.initials: C
authors.lastname: López-Otín
authors.forename: R
authors.initials: R
authors.lastname: Rodríguez
mesh_terms: D055516:2S Albumins, Plant; D000418:Albumins / Q000737:chemistry*; D000595:Amino Acid Sequence; D001937:Brassica; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry*; D011487:Protein Conformation; D013696:Temperature
chemical_list: D055516:2S Albumins, Plant; D000418:Albumins; D010940:Plant Proteins; C050363:napin protein, Brassica napus