title: Molecular cloning, isolation, and properties of chaperone Skp from Yersinia pseudotuberculosis.
abstract: The skp gene of Yersinia pseudotuberculosis was expressed without its signal sequence in Escherichia coli BL21(DE3) cells. The recombinant protein Skp accumulated in soluble form in the cytoplasm of the producer strain. The protein was isolated and characterized: the molecular weight, isoelectric point, N-terminal amino acid sequence (20 amino acid residues), and the content of the secondary structure elements were determined. Using cross-linking stabilization and high-mass MALDI-TOF mass spectrometric analysis, it was found that rSkp forms a stable homotrimer in solution and interacts with human IgG. Three-dimensional models of the Skp trimer and its complexes with Fc- and Fab-fragments of human IgG1 were constructed by computer modeling.
authors.affiliation: Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences, Vladivostok, 690022, Russia. sev1972@mail.ru
authors.forename: E V
authors.initials: EV
authors.lastname: Sidorin
authors.forename: N M
authors.initials: NM
authors.lastname: Tishchenko
authors.forename: V A
authors.initials: VA
authors.lastname: Khomenko
authors.forename: M P
authors.initials: MP
authors.lastname: Isaeva
authors.forename: P S
authors.initials: PS
authors.lastname: Dmitrenok
authors.forename: N Yu
authors.initials: NY
authors.lastname: Kim
authors.forename: G N
authors.initials: GN
authors.lastname: Likhatskaya
authors.forename: T F
authors.initials: TF
authors.lastname: Solov'eva
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002942:Circular Dichroism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D018832:Molecular Chaperones / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D016415:Sequence Alignment; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D015011:Yersinia pseudotuberculosis / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D007074:Immunoglobulin G; D018832:Molecular Chaperones; D011994:Recombinant Proteins