Difference: TwodxYjbB (1 vs. 9)

Revision 903 Apr 2014 - Main.NicolasCoudray

Changed:
<
<
META TOPICPARENT name="2DCrystallizationExp"
>
>
META TOPICPARENT name="TwoDCrystallizationGroup"
 

2dx - YjbB 2D-crystallization

Properties

MLTLLHLLSAVALLVWGTHIVRTGVMRVFGARLRTVLSRSVEKKPLAFCAGIGVTALVQSSNATTMLVTSFVAQDLVALAPALVIVLGADVGTALMARILTFDLSWLSPLLIFIGVIFFLGRKQSRAGQLGRVGIGLGLI LLALELIVQAVTPITQANGVQVIFASLTGDILLDALIGAMFAIISYSSLAAVLLTATLTAAGIISFPVALCLVIGANLGSGLLAMLNNSAANAAARRVALGSLLFKLVGSLIILPFVHLLAETMGKLSLPKAELVIYFHVFYNLVRCLVMLPFVDPMARFCKTIIRDEPELDTQLRPKHLDVSALDTPTLALANAARETLRIGDAMEQMMEGLNKVMHGEPRQEKELRKLADDINVLYTAIKLYLARMPKEELAEEESRRWAEIIEMSLNLEQASDIVERMGSEIADKSLAARRAFSLDGLKELDALYEQLLSNLKLAMSVFFSGDVTSARRLRRSKHRFRILNRRYSHAHVDRLHQQNVQSIETSSLHLGLLGDMQRLNSLFCSVAYSVLEQPDEDEGRDEY

  • 543 amino acids
  • 59.4 kDa
  • pI ~ 7.7
  • 52 negatively charged residues
  • 53 positively charged residues
  • extinction coefficient: 29910
  • instability index: 38.35
  • alipathic index: 123.98
  • GRAVY: 0.432
  • TM helices: 8 (TMHMM)
  • amino acids in the TM domain: 181 (LPR(m/m) expected: 10-30 ~ 0.15 to 0.4 in w/w; with LPR(m/m)=24>>LPR(w/w)=0.32

X166- Protein screening

4 tubes from 2 homologs were received (corresponding to one of the two peaks of the SEC): Protein_Acquisition_Form_P3A12.doc and Protein_Acquisition_Form_P9H2.doc. LPR 0.25 to 1.25 with DMPC and DOPG in DDM screened for each of the 4 protein preparation to compare them: see matrix: X166-YjbB.xlsx.

--+++Results, X166: See image selection: GM_032911-MCMJR1.pptx.
Sheets and tube-like structures growing out from aggregates at LPR 0.25 with DMPC

X133-EM267 - Lipid screening

Protein in 0.15% DM, 150 mM NaCl, 20 mM HEPES pH 7.8 (YjbB_Protein_Acquisition_Form.doc) Conditions:
  • DMPC, DOPG, E.coli polar lipids in DM
  • LPR 0.25 to 2
  • microdialysis at 27 degrees, and MBCD (manual) at 4 degrees: 1st night incubation, then 1.2 ul for 4 mornings.
  • Buffer: pH 7.5, 20 mM Hepes, 100 mM NaCl, 5 mM NaN3
  • Final [DM] at least 1.5 mg/ml
details: YjbB_screening_july2012.xlsx.

Comments (August 2012)

Image selection: X133_YjbB.ppt.
  • no gel filtration was done (it aggregated).
    • Ask him if he tried with DDM and what happened after the protein was concentrated.

  • DMPC and DOPG from LPR=1.0 (aggregates up to 0.75)
  • very bad with E.coli (Aggregates)

  • Set ALLOWTOPICVIEW =

-- NicolasCoudray - 27 Jul 2012

Added:
>
>
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1363366883" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115197" name="Protein_Acquisition_Form_P9H2.doc" path="Protein_Acquisition_Form_P9H2.doc" size="61440" user="Main.NicolasCoudray" version="1"
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115182" name="X166-YjbB.xlsx" path="X166-YjbB.xlsx" size="668563" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115191" name="Protein_Acquisition_Form_P3A12.doc" path="Protein_Acquisition_Form_P3A12.doc" size="61440" user="Main.NicolasCoudray" version="1"
Deleted:
<
<
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1363366883" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" user="Main.NicolasCoudray" version="1"
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1343398249" name="YjbB_screening_july2012.xlsx" path="YjbB_screening_july2012.xlsx" size="31844" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1343398258" name="YjbB_Protein_Acquisition_Form.doc" path="YjbB_Protein_Acquisition_Form.doc" size="63488" user="Main.NicolasCoudray" version="1"
Deleted:
<
<
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115197" name="Protein_Acquisition_Form_P9H2.doc" path="Protein_Acquisition_Form_P9H2.doc" size="61440" user="Main.NicolasCoudray" version="1"
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1388087465" name="2013-12-26-Status_of_the_targets.pptx" path="2013-12-26-Status_of_the_targets.pptx" size="5793736" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1345574482" name="X133_YjbB.ppt" path="X133_YjbB.ppt" size="9049088" user="Main.RalphLasala" version="1"
META TOPICMOVED by="NicolasCoudray" date="1344355491" from="Main.TwodxUMAYjbB" to="Main.TwodxYjbB"

Revision 826 Dec 2013 - Main.NicolasCoudray

 
META TOPICPARENT name="2DCrystallizationExp"

2dx - YjbB 2D-crystallization

Properties

MLTLLHLLSAVALLVWGTHIVRTGVMRVFGARLRTVLSRSVEKKPLAFCAGIGVTALVQSSNATTMLVTSFVAQDLVALAPALVIVLGADVGTALMARILTFDLSWLSPLLIFIGVIFFLGRKQSRAGQLGRVGIGLGLI LLALELIVQAVTPITQANGVQVIFASLTGDILLDALIGAMFAIISYSSLAAVLLTATLTAAGIISFPVALCLVIGANLGSGLLAMLNNSAANAAARRVALGSLLFKLVGSLIILPFVHLLAETMGKLSLPKAELVIYFHVFYNLVRCLVMLPFVDPMARFCKTIIRDEPELDTQLRPKHLDVSALDTPTLALANAARETLRIGDAMEQMMEGLNKVMHGEPRQEKELRKLADDINVLYTAIKLYLARMPKEELAEEESRRWAEIIEMSLNLEQASDIVERMGSEIADKSLAARRAFSLDGLKELDALYEQLLSNLKLAMSVFFSGDVTSARRLRRSKHRFRILNRRYSHAHVDRLHQQNVQSIETSSLHLGLLGDMQRLNSLFCSVAYSVLEQPDEDEGRDEY

  • 543 amino acids
  • 59.4 kDa
  • pI ~ 7.7
  • 52 negatively charged residues
  • 53 positively charged residues
  • extinction coefficient: 29910
  • instability index: 38.35
  • alipathic index: 123.98
  • GRAVY: 0.432
  • TM helices: 8 (TMHMM)
  • amino acids in the TM domain: 181 (LPR(m/m) expected: 10-30 ~ 0.15 to 0.4 in w/w; with LPR(m/m)=24>>LPR(w/w)=0.32

X166- Protein screening

4 tubes from 2 homologs were received (corresponding to one of the two peaks of the SEC): Protein_Acquisition_Form_P3A12.doc and Protein_Acquisition_Form_P9H2.doc. LPR 0.25 to 1.25 with DMPC and DOPG in DDM screened for each of the 4 protein preparation to compare them: see matrix: X166-YjbB.xlsx.

--+++Results, X166: See image selection: GM_032911-MCMJR1.pptx.
Sheets and tube-like structures growing out from aggregates at LPR 0.25 with DMPC

X133-EM267 - Lipid screening

Protein in 0.15% DM, 150 mM NaCl, 20 mM HEPES pH 7.8 (YjbB_Protein_Acquisition_Form.doc) Conditions:
  • DMPC, DOPG, E.coli polar lipids in DM
  • LPR 0.25 to 2
  • microdialysis at 27 degrees, and MBCD (manual) at 4 degrees: 1st night incubation, then 1.2 ul for 4 mornings.
  • Buffer: pH 7.5, 20 mM Hepes, 100 mM NaCl, 5 mM NaN3
  • Final [DM] at least 1.5 mg/ml
details: YjbB_screening_july2012.xlsx.

Comments (August 2012)

Image selection: X133_YjbB.ppt.
  • no gel filtration was done (it aggregated).
    • Ask him if he tried with DDM and what happened after the protein was concentrated.

  • DMPC and DOPG from LPR=1.0 (aggregates up to 0.75)
  • very bad with E.coli (Aggregates)

  • Set ALLOWTOPICVIEW =

-- NicolasCoudray - 27 Jul 2012

Added:
>
>
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115182" name="X166-YjbB.xlsx" path="X166-YjbB.xlsx" size="668563" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115191" name="Protein_Acquisition_Form_P3A12.doc" path="Protein_Acquisition_Form_P3A12.doc" size="61440" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1363366883" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1343398249" name="YjbB_screening_july2012.xlsx" path="YjbB_screening_july2012.xlsx" size="31844" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1343398258" name="YjbB_Protein_Acquisition_Form.doc" path="YjbB_Protein_Acquisition_Form.doc" size="63488" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115197" name="Protein_Acquisition_Form_P9H2.doc" path="Protein_Acquisition_Form_P9H2.doc" size="61440" user="Main.NicolasCoudray" version="1"
Added:
>
>
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1388087465" name="2013-12-26-Status_of_the_targets.pptx" path="2013-12-26-Status_of_the_targets.pptx" size="5793736" user="Main.NicolasCoudray" version="1"
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1345574482" name="X133_YjbB.ppt" path="X133_YjbB.ppt" size="9049088" user="Main.RalphLasala" version="1"
META TOPICMOVED by="NicolasCoudray" date="1344355491" from="Main.TwodxUMAYjbB" to="Main.TwodxYjbB"

Revision 715 Mar 2013 - Main.NicolasCoudray

 
META TOPICPARENT name="2DCrystallizationExp"

2dx - YjbB 2D-crystallization

Properties

MLTLLHLLSAVALLVWGTHIVRTGVMRVFGARLRTVLSRSVEKKPLAFCAGIGVTALVQSSNATTMLVTSFVAQDLVALAPALVIVLGADVGTALMARILTFDLSWLSPLLIFIGVIFFLGRKQSRAGQLGRVGIGLGLI LLALELIVQAVTPITQANGVQVIFASLTGDILLDALIGAMFAIISYSSLAAVLLTATLTAAGIISFPVALCLVIGANLGSGLLAMLNNSAANAAARRVALGSLLFKLVGSLIILPFVHLLAETMGKLSLPKAELVIYFHVFYNLVRCLVMLPFVDPMARFCKTIIRDEPELDTQLRPKHLDVSALDTPTLALANAARETLRIGDAMEQMMEGLNKVMHGEPRQEKELRKLADDINVLYTAIKLYLARMPKEELAEEESRRWAEIIEMSLNLEQASDIVERMGSEIADKSLAARRAFSLDGLKELDALYEQLLSNLKLAMSVFFSGDVTSARRLRRSKHRFRILNRRYSHAHVDRLHQQNVQSIETSSLHLGLLGDMQRLNSLFCSVAYSVLEQPDEDEGRDEY

  • 543 amino acids
  • 59.4 kDa
  • pI ~ 7.7
  • 52 negatively charged residues
  • 53 positively charged residues
  • extinction coefficient: 29910
  • instability index: 38.35
  • alipathic index: 123.98
  • GRAVY: 0.432
  • TM helices: 8 (TMHMM)
  • amino acids in the TM domain: 181 (LPR(m/m) expected: 10-30 ~ 0.15 to 0.4 in w/w; with LPR(m/m)=24>>LPR(w/w)=0.32

X166- Protein screening

4 tubes from 2 homologs were received (corresponding to one of the two peaks of the SEC): Protein_Acquisition_Form_P3A12.doc and Protein_Acquisition_Form_P9H2.doc. LPR 0.25 to 1.25 with DMPC and DOPG in DDM screened for each of the 4 protein preparation to compare them: see matrix: X166-YjbB.xlsx.

Added:
>
>		--+++Results, X166: See image selection: GM_032911-MCMJR1.pptx.
Sheets and tube-like structures growing out from aggregates at LPR 0.25 with DMPC
 
Added:
>
>
 

X133-EM267 - Lipid screening

Protein in 0.15% DM, 150 mM NaCl, 20 mM HEPES pH 7.8 (YjbB_Protein_Acquisition_Form.doc) Conditions:
  • DMPC, DOPG, E.coli polar lipids in DM
  • LPR 0.25 to 2
  • microdialysis at 27 degrees, and MBCD (manual) at 4 degrees: 1st night incubation, then 1.2 ul for 4 mornings.
  • Buffer: pH 7.5, 20 mM Hepes, 100 mM NaCl, 5 mM NaN3
  • Final [DM] at least 1.5 mg/ml
details: YjbB_screening_july2012.xlsx.

Comments (August 2012)

Image selection: X133_YjbB.ppt.
  • no gel filtration was done (it aggregated).
    • Ask him if he tried with DDM and what happened after the protein was concentrated.

  • DMPC and DOPG from LPR=1.0 (aggregates up to 0.75)
  • very bad with E.coli (Aggregates)

  • Set ALLOWTOPICVIEW =

-- NicolasCoudray - 27 Jul 2012

Deleted:
<
<		 * GM_032911-MCMJR1.pptx: GM_032911-MCMJR1.pptx d69 1
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115182" name="X166-YjbB.xlsx" path="X166-YjbB.xlsx" size="668563" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115191" name="Protein_Acquisition_Form_P3A12.doc" path="Protein_Acquisition_Form_P3A12.doc" size="61440" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1363366883" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1343398249" name="YjbB_screening_july2012.xlsx" path="YjbB_screening_july2012.xlsx" size="31844" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1343398258" name="YjbB_Protein_Acquisition_Form.doc" path="YjbB_Protein_Acquisition_Form.doc" size="63488" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115197" name="Protein_Acquisition_Form_P9H2.doc" path="Protein_Acquisition_Form_P9H2.doc" size="61440" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1345574482" name="X133_YjbB.ppt" path="X133_YjbB.ppt" size="9049088" user="Main.RalphLasala" version="1"
Deleted:
<
<
META FILEATTACHMENT attachment="GM_032911-MCMJR1.pptx" attr="" comment="" date="1363366882" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" stream="GM_032911-MCMJR1.pptx" user="Main.NicolasCoudray" version="1"
 
META TOPICMOVED by="NicolasCoudray" date="1344355491" from="Main.TwodxUMAYjbB" to="Main.TwodxYjbB"

Revision 615 Mar 2013 - Main.NicolasCoudray

 
META TOPICPARENT name="2DCrystallizationExp"

2dx - YjbB 2D-crystallization

Properties

MLTLLHLLSAVALLVWGTHIVRTGVMRVFGARLRTVLSRSVEKKPLAFCAGIGVTALVQSSNATTMLVTSFVAQDLVALAPALVIVLGADVGTALMARILTFDLSWLSPLLIFIGVIFFLGRKQSRAGQLGRVGIGLGLI LLALELIVQAVTPITQANGVQVIFASLTGDILLDALIGAMFAIISYSSLAAVLLTATLTAAGIISFPVALCLVIGANLGSGLLAMLNNSAANAAARRVALGSLLFKLVGSLIILPFVHLLAETMGKLSLPKAELVIYFHVFYNLVRCLVMLPFVDPMARFCKTIIRDEPELDTQLRPKHLDVSALDTPTLALANAARETLRIGDAMEQMMEGLNKVMHGEPRQEKELRKLADDINVLYTAIKLYLARMPKEELAEEESRRWAEIIEMSLNLEQASDIVERMGSEIADKSLAARRAFSLDGLKELDALYEQLLSNLKLAMSVFFSGDVTSARRLRRSKHRFRILNRRYSHAHVDRLHQQNVQSIETSSLHLGLLGDMQRLNSLFCSVAYSVLEQPDEDEGRDEY

  • 543 amino acids
  • 59.4 kDa
  • pI ~ 7.7
  • 52 negatively charged residues
  • 53 positively charged residues
  • extinction coefficient: 29910
  • instability index: 38.35
  • alipathic index: 123.98
  • GRAVY: 0.432
  • TM helices: 8 (TMHMM)
  • amino acids in the TM domain: 181 (LPR(m/m) expected: 10-30 ~ 0.15 to 0.4 in w/w; with LPR(m/m)=24>>LPR(w/w)=0.32

X166- Protein screening

4 tubes from 2 homologs were received (corresponding to one of the two peaks of the SEC): Protein_Acquisition_Form_P3A12.doc and Protein_Acquisition_Form_P9H2.doc. LPR 0.25 to 1.25 with DMPC and DOPG in DDM screened for each of the 4 protein preparation to compare them: see matrix: X166-YjbB.xlsx.

X133-EM267 - Lipid screening

Protein in 0.15% DM, 150 mM NaCl, 20 mM HEPES pH 7.8 (YjbB_Protein_Acquisition_Form.doc) Conditions:
  • DMPC, DOPG, E.coli polar lipids in DM
  • LPR 0.25 to 2
  • microdialysis at 27 degrees, and MBCD (manual) at 4 degrees: 1st night incubation, then 1.2 ul for 4 mornings.
  • Buffer: pH 7.5, 20 mM Hepes, 100 mM NaCl, 5 mM NaN3
  • Final [DM] at least 1.5 mg/ml
details: YjbB_screening_july2012.xlsx.

Comments (August 2012)

Image selection: X133_YjbB.ppt.
  • no gel filtration was done (it aggregated).
    • Ask him if he tried with DDM and what happened after the protein was concentrated.

  • DMPC and DOPG from LPR=1.0 (aggregates up to 0.75)
  • very bad with E.coli (Aggregates)

  • Set ALLOWTOPICVIEW =

-- NicolasCoudray - 27 Jul 2012

Added:
>
>		 * GM_032911-MCMJR1.pptx: GM_032911-MCMJR1.pptx d69 1
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115182" name="X166-YjbB.xlsx" path="X166-YjbB.xlsx" size="668563" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115191" name="Protein_Acquisition_Form_P3A12.doc" path="Protein_Acquisition_Form_P3A12.doc" size="61440" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1363366883" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1343398249" name="YjbB_screening_july2012.xlsx" path="YjbB_screening_july2012.xlsx" size="31844" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1343398258" name="YjbB_Protein_Acquisition_Form.doc" path="YjbB_Protein_Acquisition_Form.doc" size="63488" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115197" name="Protein_Acquisition_Form_P9H2.doc" path="Protein_Acquisition_Form_P9H2.doc" size="61440" user="Main.NicolasCoudray" version="1"
Added:
>
>
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1345574482" name="X133_YjbB.ppt" path="X133_YjbB.ppt" size="9049088" user="Main.RalphLasala" version="1"
 
META FILEATTACHMENT attachment="GM_032911-MCMJR1.pptx" attr="" comment="" date="1363366882" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" stream="GM_032911-MCMJR1.pptx" user="Main.NicolasCoudray" version="1"
META TOPICMOVED by="NicolasCoudray" date="1344355491" from="Main.TwodxUMAYjbB" to="Main.TwodxYjbB"

Revision 521 Dec 2012 - Main.NicolasCoudray

 
META TOPICPARENT name="2DCrystallizationExp"

2dx - YjbB 2D-crystallization

Properties

MLTLLHLLSAVALLVWGTHIVRTGVMRVFGARLRTVLSRSVEKKPLAFCAGIGVTALVQSSNATTMLVTSFVAQDLVALAPALVIVLGADVGTALMARILTFDLSWLSPLLIFIGVIFFLGRKQSRAGQLGRVGIGLGLI LLALELIVQAVTPITQANGVQVIFASLTGDILLDALIGAMFAIISYSSLAAVLLTATLTAAGIISFPVALCLVIGANLGSGLLAMLNNSAANAAARRVALGSLLFKLVGSLIILPFVHLLAETMGKLSLPKAELVIYFHVFYNLVRCLVMLPFVDPMARFCKTIIRDEPELDTQLRPKHLDVSALDTPTLALANAARETLRIGDAMEQMMEGLNKVMHGEPRQEKELRKLADDINVLYTAIKLYLARMPKEELAEEESRRWAEIIEMSLNLEQASDIVERMGSEIADKSLAARRAFSLDGLKELDALYEQLLSNLKLAMSVFFSGDVTSARRLRRSKHRFRILNRRYSHAHVDRLHQQNVQSIETSSLHLGLLGDMQRLNSLFCSVAYSVLEQPDEDEGRDEY

  • 543 amino acids
  • 59.4 kDa
  • pI ~ 7.7
  • 52 negatively charged residues
  • 53 positively charged residues
  • extinction coefficient: 29910
  • instability index: 38.35
  • alipathic index: 123.98
  • GRAVY: 0.432
  • TM helices: 8 (TMHMM)
  • amino acids in the TM domain: 181 (LPR(m/m) expected: 10-30 ~ 0.15 to 0.4 in w/w; with LPR(m/m)=24>>LPR(w/w)=0.32
Added:
>
>

X166- Protein screening

4 tubes from 2 homologs were received (corresponding to one of the two peaks of the SEC): Protein_Acquisition_Form_P3A12.doc and Protein_Acquisition_Form_P9H2.doc. LPR 0.25 to 1.25 with DMPC and DOPG in DDM screened for each of the 4 protein preparation to compare them: see matrix: X166-YjbB.xlsx.
 
Added:
>
>

 

X133-EM267 - Lipid screening

Protein in 0.15% DM, 150 mM NaCl, 20 mM HEPES pH 7.8 (YjbB_Protein_Acquisition_Form.doc) Conditions:
  • DMPC, DOPG, E.coli polar lipids in DM
  • LPR 0.25 to 2
  • microdialysis at 27 degrees, and MBCD (manual) at 4 degrees: 1st night incubation, then 1.2 ul for 4 mornings.
  • Buffer: pH 7.5, 20 mM Hepes, 100 mM NaCl, 5 mM NaN3
  • Final [DM] at least 1.5 mg/ml
details: YjbB_screening_july2012.xlsx.

Comments (August 2012)

Image selection: X133_YjbB.ppt.
  • no gel filtration was done (it aggregated).
    • Ask him if he tried with DDM and what happened after the protein was concentrated.

  • DMPC and DOPG from LPR=1.0 (aggregates up to 0.75)
  • very bad with E.coli (Aggregates)

  • Set ALLOWTOPICVIEW =

-- NicolasCoudray - 27 Jul 2012

Added:
>
>
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115182" name="X166-YjbB.xlsx" path="X166-YjbB.xlsx" size="668563" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115191" name="Protein_Acquisition_Form_P3A12.doc" path="Protein_Acquisition_Form_P3A12.doc" size="61440" user="Main.NicolasCoudray" version="1"
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1363366883" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1343398249" name="YjbB_screening_july2012.xlsx" path="YjbB_screening_july2012.xlsx" size="31844" user="Main.NicolasCoudray" version="1"
Added:
>
>
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1343398258" name="YjbB_Protein_Acquisition_Form.doc" path="YjbB_Protein_Acquisition_Form.doc" size="63488" user="Main.NicolasCoudray" version="1"
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115197" name="Protein_Acquisition_Form_P9H2.doc" path="Protein_Acquisition_Form_P9H2.doc" size="61440" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attachment="GM_032911-MCMJR1.pptx" attr="" comment="" date="1363366882" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" stream="GM_032911-MCMJR1.pptx" user="Main.NicolasCoudray" version="1"
META TOPICMOVED by="NicolasCoudray" date="1344355491" from="Main.TwodxUMAYjbB" to="Main.TwodxYjbB"

Revision 422 Aug 2012 - Main.NicolasCoudray

 
META TOPICPARENT name="2DCrystallizationExp"

2dx - YjbB 2D-crystallization

Properties

MLTLLHLLSAVALLVWGTHIVRTGVMRVFGARLRTVLSRSVEKKPLAFCAGIGVTALVQSSNATTMLVTSFVAQDLVALAPALVIVLGADVGTALMARILTFDLSWLSPLLIFIGVIFFLGRKQSRAGQLGRVGIGLGLI LLALELIVQAVTPITQANGVQVIFASLTGDILLDALIGAMFAIISYSSLAAVLLTATLTAAGIISFPVALCLVIGANLGSGLLAMLNNSAANAAARRVALGSLLFKLVGSLIILPFVHLLAETMGKLSLPKAELVIYFHVFYNLVRCLVMLPFVDPMARFCKTIIRDEPELDTQLRPKHLDVSALDTPTLALANAARETLRIGDAMEQMMEGLNKVMHGEPRQEKELRKLADDINVLYTAIKLYLARMPKEELAEEESRRWAEIIEMSLNLEQASDIVERMGSEIADKSLAARRAFSLDGLKELDALYEQLLSNLKLAMSVFFSGDVTSARRLRRSKHRFRILNRRYSHAHVDRLHQQNVQSIETSSLHLGLLGDMQRLNSLFCSVAYSVLEQPDEDEGRDEY

  • 543 amino acids
  • 59.4 kDa
  • pI ~ 7.7
  • 52 negatively charged residues
  • 53 positively charged residues
  • extinction coefficient: 29910
  • instability index: 38.35
  • alipathic index: 123.98
  • GRAVY: 0.432
  • TM helices: 8 (TMHMM)
  • amino acids in the TM domain: 181 (LPR(m/m) expected: 10-30 ~ 0.15 to 0.4 in w/w; with LPR(m/m)=24>>LPR(w/w)=0.32

X133-EM267 - Lipid screening

Protein in 0.15% DM, 150 mM NaCl, 20 mM HEPES pH 7.8 (YjbB_Protein_Acquisition_Form.doc) Conditions:
  • DMPC, DOPG, E.coli polar lipids in DM
  • LPR 0.25 to 2
  • microdialysis at 27 degrees, and MBCD (manual) at 4 degrees: 1st night incubation, then 1.2 ul for 4 mornings.
  • Buffer: pH 7.5, 20 mM Hepes, 100 mM NaCl, 5 mM NaN3
  • Final [DM] at least 1.5 mg/ml
details: YjbB_screening_july2012.xlsx.

Comments (August 2012)

Added:
>
>		Image selection: X133_YjbB.ppt.
 
  • no gel filtration was done (it aggregated).
    • Ask him if he tried with DDM and what happened after the protein was concentrated.

  • DMPC and DOPG from LPR=1.0 (aggregates up to 0.75)
  • very bad with E.coli (Aggregates)

  • Set ALLOWTOPICVIEW =

-- NicolasCoudray - 27 Jul 2012

Deleted:
<
<		 * X133_YjbB.ppt: X133_YjbB.ppt
 
Added:
>
>
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1363366883" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1343398249" name="YjbB_screening_july2012.xlsx" path="YjbB_screening_july2012.xlsx" size="31844" user="Main.NicolasCoudray" version="1"
Changed:
<
<
META FILEATTACHMENT attachment="X133_YjbB.ppt" attr="" comment="" date="1345574481" name="X133_YjbB.ppt" path="X133_YjbB.ppt" size="9049088" stream="X133_YjbB.ppt" user="Main.RalphLasala" version="1"
>
>
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1356115197" name="Protein_Acquisition_Form_P9H2.doc" path="Protein_Acquisition_Form_P9H2.doc" size="61440" user="Main.NicolasCoudray" version="1"
 
META FILEATTACHMENT attachment="GM_032911-MCMJR1.pptx" attr="" comment="" date="1363366882" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" stream="GM_032911-MCMJR1.pptx" user="Main.NicolasCoudray" version="1"
META TOPICMOVED by="NicolasCoudray" date="1344355491" from="Main.TwodxUMAYjbB" to="Main.TwodxYjbB"

Revision 321 Aug 2012 - Main.RalphLasala

 
META TOPICPARENT name="2DCrystallizationExp"

2dx - YjbB 2D-crystallization

Properties

MLTLLHLLSAVALLVWGTHIVRTGVMRVFGARLRTVLSRSVEKKPLAFCAGIGVTALVQSSNATTMLVTSFVAQDLVALAPALVIVLGADVGTALMARILTFDLSWLSPLLIFIGVIFFLGRKQSRAGQLGRVGIGLGLI LLALELIVQAVTPITQANGVQVIFASLTGDILLDALIGAMFAIISYSSLAAVLLTATLTAAGIISFPVALCLVIGANLGSGLLAMLNNSAANAAARRVALGSLLFKLVGSLIILPFVHLLAETMGKLSLPKAELVIYFHVFYNLVRCLVMLPFVDPMARFCKTIIRDEPELDTQLRPKHLDVSALDTPTLALANAARETLRIGDAMEQMMEGLNKVMHGEPRQEKELRKLADDINVLYTAIKLYLARMPKEELAEEESRRWAEIIEMSLNLEQASDIVERMGSEIADKSLAARRAFSLDGLKELDALYEQLLSNLKLAMSVFFSGDVTSARRLRRSKHRFRILNRRYSHAHVDRLHQQNVQSIETSSLHLGLLGDMQRLNSLFCSVAYSVLEQPDEDEGRDEY

  • 543 amino acids
  • 59.4 kDa
  • pI ~ 7.7
  • 52 negatively charged residues
  • 53 positively charged residues
  • extinction coefficient: 29910
  • instability index: 38.35
  • alipathic index: 123.98
  • GRAVY: 0.432
  • TM helices: 8 (TMHMM)
  • amino acids in the TM domain: 181 (LPR(m/m) expected: 10-30 ~ 0.15 to 0.4 in w/w; with LPR(m/m)=24>>LPR(w/w)=0.32

X133-EM267 - Lipid screening

Protein in 0.15% DM, 150 mM NaCl, 20 mM HEPES pH 7.8 (YjbB_Protein_Acquisition_Form.doc) Conditions:
  • DMPC, DOPG, E.coli polar lipids in DM
  • LPR 0.25 to 2
  • microdialysis at 27 degrees, and MBCD (manual) at 4 degrees: 1st night incubation, then 1.2 ul for 4 mornings.
  • Buffer: pH 7.5, 20 mM Hepes, 100 mM NaCl, 5 mM NaN3
  • Final [DM] at least 1.5 mg/ml
details: YjbB_screening_july2012.xlsx.

Comments (August 2012)

  • no gel filtration was done (it aggregated).
    • Ask him if he tried with DDM and what happened after the protein was concentrated.

  • DMPC and DOPG from LPR=1.0 (aggregates up to 0.75)
  • very bad with E.coli (Aggregates)

  • Set ALLOWTOPICVIEW =

-- NicolasCoudray - 27 Jul 2012

Added:
>
>		 * X133_YjbB.ppt: X133_YjbB.ppt
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1363366883" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1343398249" name="YjbB_screening_july2012.xlsx" path="YjbB_screening_july2012.xlsx" size="31844" user="Main.NicolasCoudray" version="1"
Added:
>
>
META FILEATTACHMENT attachment="X133_YjbB.ppt" attr="" comment="" date="1345574481" name="X133_YjbB.ppt" path="X133_YjbB.ppt" size="9049088" stream="X133_YjbB.ppt" user="Main.RalphLasala" version="1"
 
META FILEATTACHMENT attachment="GM_032911-MCMJR1.pptx" attr="" comment="" date="1363366882" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" stream="GM_032911-MCMJR1.pptx" user="Main.NicolasCoudray" version="1"
META TOPICMOVED by="NicolasCoudray" date="1344355491" from="Main.TwodxUMAYjbB" to="Main.TwodxYjbB"

Revision 207 Aug 2012 - Main.NicolasCoudray

 
META TOPICPARENT name="2DCrystallizationExp"

2dx - YjbB 2D-crystallization

Properties

MLTLLHLLSAVALLVWGTHIVRTGVMRVFGARLRTVLSRSVEKKPLAFCAGIGVTALVQSSNATTMLVTSFVAQDLVALAPALVIVLGADVGTALMARILTFDLSWLSPLLIFIGVIFFLGRKQSRAGQLGRVGIGLGLI LLALELIVQAVTPITQANGVQVIFASLTGDILLDALIGAMFAIISYSSLAAVLLTATLTAAGIISFPVALCLVIGANLGSGLLAMLNNSAANAAARRVALGSLLFKLVGSLIILPFVHLLAETMGKLSLPKAELVIYFHVFYNLVRCLVMLPFVDPMARFCKTIIRDEPELDTQLRPKHLDVSALDTPTLALANAARETLRIGDAMEQMMEGLNKVMHGEPRQEKELRKLADDINVLYTAIKLYLARMPKEELAEEESRRWAEIIEMSLNLEQASDIVERMGSEIADKSLAARRAFSLDGLKELDALYEQLLSNLKLAMSVFFSGDVTSARRLRRSKHRFRILNRRYSHAHVDRLHQQNVQSIETSSLHLGLLGDMQRLNSLFCSVAYSVLEQPDEDEGRDEY

  • 543 amino acids
  • 59.4 kDa
  • pI ~ 7.7
  • 52 negatively charged residues
  • 53 positively charged residues
  • extinction coefficient: 29910
  • instability index: 38.35
  • alipathic index: 123.98
  • GRAVY: 0.432
  • TM helices: 8 (TMHMM)
  • amino acids in the TM domain: 181 (LPR(m/m) expected: 10-30 ~ 0.15 to 0.4 in w/w; with LPR(m/m)=24>>LPR(w/w)=0.32

X133-EM267 - Lipid screening

Protein in 0.15% DM, 150 mM NaCl, 20 mM HEPES pH 7.8 (YjbB_Protein_Acquisition_Form.doc) Conditions:
  • DMPC, DOPG, E.coli polar lipids in DM
  • LPR 0.25 to 2
  • microdialysis at 27 degrees, and MBCD (manual) at 4 degrees: 1st night incubation, then 1.2 ul for 4 mornings.
  • Buffer: pH 7.5, 20 mM Hepes, 100 mM NaCl, 5 mM NaN3
  • Final [DM] at least 1.5 mg/ml
details: YjbB_screening_july2012.xlsx.
Added:
>
>

Comments (August 2012)

  • no gel filtration was done (it aggregated).
    • Ask him if he tried with DDM and what happened after the protein was concentrated.

  • DMPC and DOPG from LPR=1.0 (aggregates up to 0.75)
  • very bad with E.coli (Aggregates)
 
  • Set ALLOWTOPICVIEW =

-- NicolasCoudray - 27 Jul 2012

META FILEATTACHMENT attr="" autoattached="1" comment="" date="1363366883" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1343398249" name="YjbB_screening_july2012.xlsx" path="YjbB_screening_july2012.xlsx" size="31844" user="Main.NicolasCoudray" version="1"
Added:
>
>
META FILEATTACHMENT attachment="GM_032911-MCMJR1.pptx" attr="" comment="" date="1363366882" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" stream="GM_032911-MCMJR1.pptx" user="Main.NicolasCoudray" version="1"
 
META TOPICMOVED by="NicolasCoudray" date="1344355491" from="Main.TwodxUMAYjbB" to="Main.TwodxYjbB"

Revision 127 Jul 2012 - Main.NicolasCoudray

 
META TOPICPARENT name="2DCrystallizationExp"

2dx - YjbB 2D-crystallization

Properties

MLTLLHLLSAVALLVWGTHIVRTGVMRVFGARLRTVLSRSVEKKPLAFCAGIGVTALVQSSNATTMLVTSFVAQDLVALAPALVIVLGADVGTALMARILTFDLSWLSPLLIFIGVIFFLGRKQSRAGQLGRVGIGLGLI LLALELIVQAVTPITQANGVQVIFASLTGDILLDALIGAMFAIISYSSLAAVLLTATLTAAGIISFPVALCLVIGANLGSGLLAMLNNSAANAAARRVALGSLLFKLVGSLIILPFVHLLAETMGKLSLPKAELVIYFHVFYNLVRCLVMLPFVDPMARFCKTIIRDEPELDTQLRPKHLDVSALDTPTLALANAARETLRIGDAMEQMMEGLNKVMHGEPRQEKELRKLADDINVLYTAIKLYLARMPKEELAEEESRRWAEIIEMSLNLEQASDIVERMGSEIADKSLAARRAFSLDGLKELDALYEQLLSNLKLAMSVFFSGDVTSARRLRRSKHRFRILNRRYSHAHVDRLHQQNVQSIETSSLHLGLLGDMQRLNSLFCSVAYSVLEQPDEDEGRDEY

  • 543 amino acids
  • 59.4 kDa
  • pI ~ 7.7
  • 52 negatively charged residues
  • 53 positively charged residues
  • extinction coefficient: 29910
  • instability index: 38.35
  • alipathic index: 123.98
  • GRAVY: 0.432
  • TM helices: 8 (TMHMM)
  • amino acids in the TM domain: 181 (LPR(m/m) expected: 10-30 ~ 0.15 to 0.4 in w/w; with LPR(m/m)=24>>LPR(w/w)=0.32

X133-EM267 - Lipid screening

Protein in 0.15% DM, 150 mM NaCl, 20 mM HEPES pH 7.8 (YjbB_Protein_Acquisition_Form.doc) Conditions:
  • DMPC, DOPG, E.coli polar lipids in DM
  • LPR 0.25 to 2
  • microdialysis at 27 degrees, and MBCD (manual) at 4 degrees: 1st night incubation, then 1.2 ul for 4 mornings.
  • Buffer: pH 7.5, 20 mM Hepes, 100 mM NaCl, 5 mM NaN3
  • Final [DM] at least 1.5 mg/ml
details: YjbB_screening_july2012.xlsx.

  • Set ALLOWTOPICVIEW =

-- NicolasCoudray - 27 Jul 2012

META FILEATTACHMENT attr="" autoattached="1" comment="" date="1363366883" name="GM_032911-MCMJR1.pptx" path="GM_032911-MCMJR1.pptx" size="5759975" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1343398249" name="YjbB_screening_july2012.xlsx" path="YjbB_screening_july2012.xlsx" size="31844" user="Main.NicolasCoudray" version="1"
META TOPICMOVED by="NicolasCoudray" date="1344355491" from="Main.TwodxUMAYjbB" to="Main.TwodxYjbB"
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