Biochemistry
Biochemistry
9th Edition
ISBN: 9781319114671
Author: Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher: W. H. Freeman
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Question

What further experiments are needed to provide confidence that the structural data obtained from the SCAM analysis reflects the structure of a properly folded AlaE?

Predicted topologies and predicted AG values: - Inside -Outside TM-helix (IN->OUT) TM-helix (OUT->IN) Signal peptide TOPCONS OCTOPUS Philius PolyPhobius SCAMPI SPOCTOPUS PDB-homology ***No homologous TM proteins detected*** 20 40 60 60 Consensus prediction (TOPCONS): TOPCONS Reliability 1 0.9 10 10 8 60 + 2 0 AG (kcal/mol) 80 100 120 140 High-resolution image 20 40 60 80 Position 100 120 140 High-resolution image.
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Transcribed Image Text:Predicted topologies and predicted AG values: - Inside -Outside TM-helix (IN->OUT) TM-helix (OUT->IN) Signal peptide TOPCONS OCTOPUS Philius PolyPhobius SCAMPI SPOCTOPUS PDB-homology ***No homologous TM proteins detected*** 20 40 60 60 Consensus prediction (TOPCONS): TOPCONS Reliability 1 0.9 10 10 8 60 + 2 0 AG (kcal/mol) 80 100 120 140 High-resolution image 20 40 60 80 Position 100 120 140 High-resolution image.
kDa 55 Cysless 35 +P 20 10 +U F16C M18C M36C +P + MTSES NEM +++mPEG5K + E39C K72C V51C + + MTSES NEM +++ MPEG5K L860 Y96C |D1066 +P MTSES NEM + + + MPEG5K G125C F139C R143C P ++ + + MTSES NEM +++ MPEG5K Fig. 1 SCAM analysis of AlaE. Western blot analysis of whole cell lysates from cells harbouring plasmids expressing cysteine-free ('Cysless') or single cysteine variants of alaE with and without treatment with NEM, MTSES and mPEG5K. PEGylated (P) and unmodified protein are indicated with arrows. Outside Membrane DeepTMHMM - Most Likely Topology | Type: alpha TM Probability Inside 0 20 40 60 80 100 120 140 1.0 0.8 0.6 0.4 0.2 DeepTMHMM - Posterior Probabilities 0.0 20 40 60 80 Sequence Membrane Inside G 100 120 Outside 140
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Transcribed Image Text:kDa 55 Cysless 35 +P 20 10 +U F16C M18C M36C +P + MTSES NEM +++mPEG5K + E39C K72C V51C + + MTSES NEM +++ MPEG5K L860 Y96C |D1066 +P MTSES NEM + + + MPEG5K G125C F139C R143C P ++ + + MTSES NEM +++ MPEG5K Fig. 1 SCAM analysis of AlaE. Western blot analysis of whole cell lysates from cells harbouring plasmids expressing cysteine-free ('Cysless') or single cysteine variants of alaE with and without treatment with NEM, MTSES and mPEG5K. PEGylated (P) and unmodified protein are indicated with arrows. Outside Membrane DeepTMHMM - Most Likely Topology | Type: alpha TM Probability Inside 0 20 40 60 80 100 120 140 1.0 0.8 0.6 0.4 0.2 DeepTMHMM - Posterior Probabilities 0.0 20 40 60 80 Sequence Membrane Inside G 100 120 Outside 140
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