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We found that the B56/BubR1 647 720 -3D complex is stable during size exclusion chromatography (SEC) (Fig. S1) and GST-pulldown (See below).
In vitro, the PPIL1·SKIP interaction is kinetically unstable and the complex disintegrates during size exclusion chromatography.
However, the complex was unstable during size exclusion chromatography (not shown), suggesting kinetic instability of the complex.
Finally, some proteins interact with the matrix during size exclusion chromatography, while SV experiments are performed in free solution in a matrix free environment.
An additional shortcoming of SAGE is that the tagging enzyme sites often occur within the genes and lead to longer tag sequences, resulting in loss of such tags during size selection.
A similar difference has previously been reported in eye-tracking during size and attractiveness judgements of single female bodies suggesting that it is a robust finding (Cornelissen, Hancock, Kiviniemi, George, & Tovée, 2009).
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However, it is also possible that disordered regions of the protein retard complex mobility during size-exclusion chromatography.
Both the DH PH domain alone and the complex of RhoA and DH PH domain appeared to be monomeric during size-exclusion chromatography.
Although the detected effects were only subtle and indicate nearly complete complex dissociation during size-exclusion chromatography, they were reproducible in several experiments.
Cul5NTD eluted at a smaller elution volume during size-exclusion chromatography (54 mL) than ASB9 EloBC (57 mL) despite having a lower molecular weight, which is likely a result of its elongated shape, causing it to behave as a larger protein by gel filtration.
AIP showed a response during both size and pattern discrimination, consistent with its activation during passive viewing, but there was no differential response between these two conditions.
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