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Figure 1. ABRF PRG2010 sample composition
Figure 2. ProteoRed members participating in the ABRF PRG2010
RESULTS
- Identify the contents of tube 1 (“I sometimes see three or four extra bands on a gel but they are weak”)
- A common set of proteins (Calcyclin-binding protein, E3 ubiquitin-protein ligase, S-phase Kinase associated protein, Ubiquitin and Catenin beta-1) have been identified by most of the participants (7 out of 10).
- Identify what is different in tube 2 that might explain why the reaction failed.
- 6 out of 10 labs found out that E3 ubiquitin-ligase SIAH1 was present in samples 1 and 3 but absent in sample 2, thus explaining the lack of activity seen in sample 2.
- In addition, 4 (+1 in the wrong sample) labs identified an E.coli protein (metal-binding protein yodA), present in sample 2. Only 2 labs hypothesized (probably correctly) the presence of yodA as a consequence of its Ni-binding properties, relating them with the poliHis tag of SIAH1.
- Identify what is restored in tube 3 that might explain why the reaction is functional again.
- 6 out of 10 labs found out that E3 ubiquitin-ligase SIAH1 was present in samples 1 and 3 but absent in sample 2, thus explaining the lack of activity seen in sample 2.
- In addition, only 2 labs hypothesized (probably correctly) that the shift found in sample 3 with respect to sample 1 corresponds to the the presence of 15N in sample 3 (about 350 atoms).
- What is the nature of the unusual doublet that is present only in tube 1, but does not seem to be related to function?
- Only 1 lab hypothesized that the doublet could correspond to the native and recombinant forms of CTNB1_HUMAN.
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