| Products/Services Used | Details | Operation |
|---|---|---|
| Peptide Synthesis | The t remaining amount of the FR dipeptide eluates were precipitated with 1:3:4 v/v chloroform:methanol:water, and precipitates were washed with methanol, dried and resuspended in 1 x Laemmli SDS sample buffer for separation by SDS-PAGE, transfer to PVDF membrane followed by immunodetection using an anti-GFP antibody (GenScript) and detection by chemiluminescence using standard procedures. | Get A Quote |
The prokaryotic N-degron pathway depends on the Clp chaperone-protease system and the ClpS adaptor for recognition of N-degron bearing substrates. Plant chloroplasts contain a diversified Clp protease, including the ClpS homolog ClpS1. Several candidate ClpS1 substrates have been identified, but the N-degron specificity is unclear. Here, we employed in vitro ClpS1 affinity assays using eight N-degron green fluorescence protein reporters containing either F, Y, L, W, I, or R in the N-terminal position. This demonstrated that ClpS1 has a restricted N-degron specificity, recognizing proteins bearing an N-terminal F or W, only weakly recognizing L, but not recognizing Y or I. This affinity is dependent on two cons... More
