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

--+++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

• 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

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

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

--+++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

• 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

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

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

X133-EM267 - Lipid screening

• 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

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

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

X133-EM267 - Lipid screening

• 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

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

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

• 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

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

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

• 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

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

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

• 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

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

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

• 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

• Set ALLOWTOPICVIEW =

-- NicolasCoudray - 27 Jul 2012

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

• 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

• Set ALLOWTOPICVIEW =

-- NicolasCoudray - 27 Jul 2012