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The SCAM data for positions V51C and Y96C are different to the other datasets. Describe how the data for these positions are different to the other positions. In each case, explain what the structural basis of these differences may be.

The SCAM data for positions V51C and Y96C are different to the other datasets. Describe how the data for these positions are different to the other positions. In each case, explain what the structural basis of these differences may be.

Biochemistry
Biochemistry
6th Edition
ISBN:9781305577206
Author:Reginald H. Garrett, Charles M. Grisham
Publisher:Reginald H. Garrett, Charles M. Grisham
Chapter31: Completing The Protein Life Cycle: Folding, Processing, And Degradation
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The SCAM data for positions V51C and Y96C are different to the other datasets. Describe how the data for these positions are different to the other positions. In each case, explain what the structural basis of these differences may be.

Outside Membrane Inside Probability 1.0 0.8 0.6 0.4 0.2 0.0 0 0 kDa 55 Cysless 35 20 F16C 20 10 20 + E39C L860 G125C + + + 40 40 +P M18C ###### + + F139C +U V51C Y96C 60 60 M36C 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. 80 K72C 80 Sequence D106C +P R143C +U ---+ MTSES + MTSES NEM + MPEG5K NEM + + + MPEG5K +P +U 100 DeepTMHMM - Most Likely Topology | Type: alpha TM 100 +P + NEM + + + MPEG5K 44U + MTSES +P + MTSES - + NEM - + + + MPEG5K DeepTMHMM- Posterior Probabilities LLIG -U 120 120 140 Membrane Inside Outside 140
Transcribed Image Text:Outside Membrane Inside Probability 1.0 0.8 0.6 0.4 0.2 0.0 0 0 kDa 55 Cysless 35 20 F16C 20 10 20 + E39C L860 G125C + + + 40 40 +P M18C ###### + + F139C +U V51C Y96C 60 60 M36C 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. 80 K72C 80 Sequence D106C +P R143C +U ---+ MTSES + MTSES NEM + MPEG5K NEM + + + MPEG5K +P +U 100 DeepTMHMM - Most Likely Topology | Type: alpha TM 100 +P + NEM + + + MPEG5K 44U + MTSES +P + MTSES - + NEM - + + + MPEG5K DeepTMHMM- Posterior Probabilities LLIG -U 120 120 140 Membrane Inside Outside 140
Predicted topologies and predicted AG values: TOPCONS OCTOPUS Philius PolyPhobius SCAMPI SPOCTOPUS PDB-homology ***No homologous TM proteins detected*** TOPCONS - Inside - Outside TM-helix (IN->OUT) TM-helix (OUT->IN) Signal peptide Consensus prediction (TOPCONS): Reliability 0.9 20 20 40 40 60 60 80 80 Position 100 100 120 120 140 10 140 8 6 2 0 AG (kcal/mol) High-resolution image High-resolution image
Transcribed Image Text:Predicted topologies and predicted AG values: TOPCONS OCTOPUS Philius PolyPhobius SCAMPI SPOCTOPUS PDB-homology ***No homologous TM proteins detected*** TOPCONS - Inside - Outside TM-helix (IN->OUT) TM-helix (OUT->IN) Signal peptide Consensus prediction (TOPCONS): Reliability 0.9 20 20 40 40 60 60 80 80 Position 100 100 120 120 140 10 140 8 6 2 0 AG (kcal/mol) High-resolution image High-resolution image
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