title: Expression of Pisum sativum SAD polypeptides in production hosts and in planta: tetrameric organization of the protein.
abstract: In Pisum sativum, the short-chain alcohol dehydrogenase-like protein (SAD) gene family consists of at least three members (SAD-A, -B, and -C). Expression of two of these genes (SAD-A and -C) in Escherichia coli or Pichia pastoris resulted in full-length soluble proteins. Purified SAD-A was used as antigen for antibody production in rabbits. With these antibodies the recombinant SAD-C protein (which was most highly expressed of the two isoforms) was shown to be a tetramer consisting of a dimer of dimers. The SAD genes are transiently expressed in plants by short exposures to ultraviolet-B radiation (UV-B), as judged by northern blotting. In turn, mRNA accumulation leads to formation of SAD protein in leaf and stem tissue upon prolonged UV-B irradiation.
authors.affiliation: Department of Science and Orebro Life Science Center, Orebro University, SE-701 82 Orebro, Sweden.
authors.forename: Nikolai
authors.initials: N
authors.lastname: Scherbak
authors.forename: Mikael
authors.initials: M
authors.lastname: Brosché
authors.forename: Anneli
authors.initials: A
authors.lastname: Ala-Häivälä
authors.forename: Hilja
authors.initials: H
authors.lastname: Strid
authors.forename: Annika
authors.initials: A
authors.lastname: Ohrfelt
authors.forename: Fredrik
authors.initials: F
authors.lastname: Nilsson
authors.forename: Ake
authors.initials: A
authors.lastname: Strid
mesh_terms: D000426:Alcohol Dehydrogenase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D015152:Blotting, Northern; D015153:Blotting, Western; D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D015870:Gene Expression; D017343:Genes, Plant; D018532:Pisum sativum / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D020836:Protein Structure, Quaternary; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014466:Ultraviolet Rays
chemical_list: D010940:Plant Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D000426:Alcohol Dehydrogenase