title: Procedure for Purification of Recombinant preMsk1p from E. coli Determines Its Properties as a Factor of tRNA Import into Yeast Mitochondria.
abstract: Mitochondrial genomes of many eukaryotic organisms do not code for the full tRNA set necessary for organellar translation. Missing tRNA species are imported from the cytosol. In particular, one out of two cytosolic lysine tRNAs of the yeast Saccharomyces cerevisiae is partially internalized by mitochondria. The key protein factor of this process is the precursor of mitochondrial lysyl-tRNA synthetase, preMsk1p. In this work, we show that recombinant preMsk1p purified from E. coli in native conditions, when used in an in vitro tRNA import system, demonstrates some properties different from those shown by the renatured protein purified from E. coli in the denatured state. We also discuss the possible mechanistic reasons for this phenomenon.
authors.affiliation: Lomonosov Moscow State University, Faculty of Biology, Moscow, 119991, Russia. kamenski_pa@mail.bio.msu.ru.
authors.forename: E V
authors.initials: EV
authors.lastname: Smirnova
authors.forename: I V
authors.initials: IV
authors.lastname: Chicherin
authors.forename: M V
authors.initials: MV
authors.lastname: Baleva
authors.forename: N S
authors.initials: NS
authors.lastname: Entelis
authors.forename: I A
authors.initials: IA
authors.lastname: Tarassov
authors.forename: P A
authors.initials: PA
authors.lastname: Kamenski
mesh_terms: D001693:Biological Transport, Active; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008250:Lysine-tRNA Ligase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D008928:Mitochondria / Q000737:chemistry / Q000235:genetics; D024101:Mitochondrial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012331:RNA, Fungal / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012357:RNA, Transfer, Lys / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D024101:Mitochondrial Proteins; D012331:RNA, Fungal; D012357:RNA, Transfer, Lys; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D008250:Lysine-tRNA Ligase