Thu, 21 Aug 2008 14:00:49 BST CiteULike: neils's Cambillau http://www.citeulike.org/user/neils/author/Cambillau CiteULike.org en-gb Copyright © 2004-2008 citeulike.org http://www.citeulike.org/user/neils/article/2319613 <i>Nat Methods, Vol. 5, No. 2. (February 2008), pp. 135-146.</i><br /><br />In selecting a method to produce a recombinant protein, a researcher is faced with a bewildering array of choices as to where to start. To facilitate decision-making, we describe a consensus 'what to try first' strategy based on our collective analysis of the expression and purification of over 10,000 different proteins. This review presents methods that could be applied at the outset of any project, a prioritized list of alternate strategies and a list of pitfalls that trip many new investigators. Protein production and purification. S Gräslund P Nordlund J Weigelt J Bray O Gileadi S Knapp U Oppermann C Arrowsmith R Hui J Ming S Dhe-Paganon HW Park A Savchenko A Yee A Edwards R Vincentelli C Cambillau R Kim SH Kim Z Rao Y Shi TC Terwilliger CY Kim LW Hung GS Waldo Y Peleg S Albeck T Unger O Dym J Prilusky JL Sussman RC Stevens SA Lesley IA Wilson A Joachimiak F Collart I Dementieva MI Donnelly WH Eschenfeldt Y Kim L Stols R Wu M Zhou SK Burley JS Emtage JM Sauder D Thompson K Bain J Luz T Gheyi F Zhang S Atwell SC Almo JB Bonanno A Fiser S Swaminathan FW Studier MR Chance A Sali TB Acton R Xiao L Zhao LC Ma JF Hunt L Tong K Cunningham M Inouye S Anderson H Janjua R Shastry CK Ho D Wang H Wang M Jiang GT Montelione DI Stuart RJ Owens S Daenke A Schütz U Heinemann S Yokoyama K Büssow KC Gunsalus doi:10.1038/nmeth.f.202 Nat Methods, Vol. 5, No. 2. (February 2008), pp. 135-146. 2008-02-01T14:36:35-00:00 2008 Nat Methods 1548-7105 5 2 135 146 expression genomics protein purification structural-genomics structure http://www.citeulike.org/user/neils/article/2287221 <i>J Biol Chem, Vol. 275, No. 42. (20 October 2000), pp. 32383-32386.</i><br /><br />While most organisms grow at temperatures ranging between 20 and 50 degrees C, many archaea and a few bacteria have been found capable of withstanding temperatures close to 100 degrees C, or beyond, such as Pyrococcus or Aquifex. Here we report the results of two independent large scale unbiased approaches to identify global protein properties correlating with an extreme thermophile lifestyle. First, we performed a comparative proteome analyses using 30 complete genome sequences from the three kingdoms. A large difference between the proportions of charged versus polar (noncharged) amino acids was found to be a signature of all hyperthermophilic organisms. Second, we analyzed the water accessible surfaces of 189 protein structures belonging to mesophiles or hyperthermophiles. We found that the surfaces of hyperthermophilic proteins exhibited the shift already observed at the genomic level, i.e. a proportion of solvent accessible charged residues strongly increased at the expense of polar residues. The biophysical requirements for the presence of charged residues at the protein surface, allowing protein stabilization through ion bonds, is therefore clearly imprinted and detectable in all genome sequences available to date. Structural and genomic correlates of hyperthermostability. C Cambillau JM Claverie doi:10.1074/jbc.C000497200 J Biol Chem, Vol. 275, No. 42. (20 October 2000), pp. 32383-32386. 2008-01-25T06:19:24-00:00 2000 J Biol Chem 0021-9258 275 42 32383 32386 for-thuber genomics hyperthermophily structure thermal