2dx - EmrE (Shewanella Oneidensis) 2D-crystallization trials

Properties

• Amino acid sequence:

M S W L L L I L A G L F E I G W A I G L K Y T D G F T R L W P S V F T V L S M A L S V L L L G L A V K Q L P I G T A Y G V W V G I G A M G T A I A G I I L L G E G V S L L K I A S L I L I L L G V L G L K L A H A E N L Y F Q G

• 112 amino acids
• 11.8 kDa, Expasy (10.9 kDa, Nanda)
• pI ~ 8.09
• 4 negatively charged residues
• 5 positively charged residues
• extinction coefficient: 26470
• instability index: 19.76
• alipathic index: 159.38
• GRAVY: 1.298
• TM helices: 4
• amino acids in the TM domain: 82 (LPR(m/m) expected: 1-40 ~ 0.03 to 1.4 in w/w; with LPR(m/m)=24>>LPR(w/w)=0.68; LPR(m/m)=5>>LPR(w/w)=0.14; LPR(m/m)=40>>LPR(w/w)=1.12;)

Experiment of December 2011 (X94)

• 5 Lipids in 5 detergent screening
• Buffer: pH8, 20mM sodium phosphate, 100mM NaCl^?. 2mM MgCl^?, 1mM EDTA, 4mM DTT, 4mM mercapto-ethanol, 5mM sodium azide.
• Temperature: 25 degrees
• microdialysis plate

Set Dec 15th.

Comments (Jan 2012)

Experiment of June 2011 (X72-EM.194)

Protocol

conditions to screen:

• Dialysis buffer: 20mM sodium phosphate, 100mM NaCl. 2mM MgCl, 1mM EDTA, 4mM DTT, 4mM mercapto-ethanol, 5mM sodium azide.
• pH: 7 and 8
• Lipids prepared in DDM and C12E8:
  • DMPC
  • E.Coli
  • POPC (C12E8 only)
• Temperature: 25 degrees
• LPR: 0.5 to 1
• dialysis block

See Protocol_EmrE_oneidensis_2011_June.xls for details.

2 weeks (Set 24th June; collected July 8th)

July 21th Meeting - comments

• All is aggregated.
• Controls of protein only and lipids only are OK
• Yiip control are bad -> do experiments to find out why...

• Set ALLOWTOPICVIEW =

-- NicolasCoudray - 23 Jun 2011