Difference: TwodxGtrB (5 vs. 6)

Revision 606 Jun 2014 - Main.NicolasCoudray

 
META TOPICPARENT name="TwoDCrystallizationGroup"

2dx - 2D-crystallization trials of GtrB

Properties

  • Amino acid sequence:
HHHHHHSSGVDLGTENLYFQSNATIELSIVIPMYNEEDNLEHLFARLLEVLTPLKITYEIICVNDGSKDKTLKQLIDCYQSNRQIKIVNLSRNFGKEIALSAGIDYAQGNAVIPIDADLQDPPELIHELVDKWREGYDIVYATRRSRQGETWVKQFTAKMFYKVIGRMTEIKIPPNTGDFRLMDRKVVNAIKQLPERTRFMKGLFAWVGYRQTFVLFDREPRFQGQTKWNYWKLWNFALDGIFSFSLLPLKVWTYLGSIISLLSLAYASFLILKTITLGVDVPGYASLMVAILFLGGVQLISLGVIGEYLGRVYEEVKARPLYLVSDLWGLEYLPLEKLN
  • 340 amino acids
  • 39.1 kDa
  • pI ~ 7.24
  • 37 negatively charged residues
  • 37 positively charged residues
  • extinction coefficient: 69330
  • instability index: 29.32
  • alipathic index: 107.21
  • GRAVY: -0.024
  • TM helices: 2 (using TMHMM)
Changed:
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X285 (June 6, 2014) - "star-like" screening

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X305 (June 6, 2014) - "star-like" screening

 Best conditions of last time (pH 8, 50mM NaCl2, 2 mM MgCl2, DMPC:DOPS 8.5:1.5), changing:
  • Lipids (Yeast, DMPC:DOPS 8.5:1.5 or 1:1, Cardiolipin:DOPC 8:2-C12E7, DMPC:DOPG 8.5:1.5, DOPC:DOPE:brain 8:1:1-DMNG
  • salt: 300 mM NaCl? + 2mM MgCl2, or 50 mM KCl + 5 mM CaCl2 or without 5mM EDTA
  • pH: 5, 7 or 9
  • 4 or 37C

Details: X305-GtrB.xlsx.

X280 (March 11, 2014) - Sparse Matrix + 3dx buffers

Protein in 20 mM Na-HEPES pH 7.0, 200 mM NaCl, 0.2% (w/v) DM, and 1 mM TCEP. Purif: GtrB_March_10_2014.docx.

Acquisition form: Protein_acquisition_form-Mar11-1_GtrB.doc.

Matrix:

  • Sparse
  • 3dx buffers @4C + lipids in DM (OG or Triton X100)
Details: X280-GtrB_Sparse.xlsx.

X280, results

Mostly vesicles, a few non-diffracting tubes.
In general, conditions with DMPC seem better (larger vesicles or tubes):
  • Some tubes @ pH8, 50 NaCl, 2 MgCl2 (DMPC:DOPS)
  • Very rare tubes @ pH7, 150 NaCl, 40 MgCl2 (DMPC:DOPC)
  • Few large vesicles (>1um) @pH9.5, 100 NaCl (DMPC:cholesterol)
  • Striation (from lipids?) @ pH7, 300 NaCl (DMPC alone)

Protein control shows some vesicles as well.

Image selection: X280-GtrB-ImageSelection.pptx.

  • Set ALLOWTOPICVIEW =

-- NicolasCoudray - 11 Mar 2014

d97 2

Added:
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>		 * X305-GtrB.xlsx: X305-GtrB.xlsx d86 1
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1394562188" name="Protein_acquisition_form-Mar11-1_GtrB.doc" path="Protein_acquisition_form-Mar11-1_GtrB.doc" size="56832" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1394552547" name="X280-GtrB_Sparse.xlsx" path="X280-GtrB_Sparse.xlsx" size="618800" user="Main.NicolasCoudray" version="1"
Changed:
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META FILEATTACHMENT attr="" autoattached="1" comment="" date="1401837583" name="X305-GtrB.xlsx" path="X305-GtrB.xlsx" size="605856" user="Main.NicolasCoudray" version="1"
>
>
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1402077433" name="X305-GtrB.xlsx" path="X305-GtrB.xlsx" size="654594" user="Main.NicolasCoudray" version="2"
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1406118280" name="X316-GtrB.xlsx" path="X316-GtrB.xlsx" size="614895" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1406118292" name="GtrB_2dx3_july_15_2014.docx" path="GtrB_2dx3_july_15_2014.docx" size="234832" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1395674610" name="X280-GtrB-ImageSelection.pptx" path="X280-GtrB-ImageSelection.pptx" size="9385477" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1394552555" name="GtrB_March_10_2014.docx" path="GtrB_March_10_2014.docx" size="269272" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1402922402" name="X305-GtrB-Summary.pptx" path="X305-GtrB-Summary.pptx" size="7364006" user="Main.NicolasCoudray" version="1"
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