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Differently from studies so far that looked at oligomeric re-organization of DegP into larger 12 and 24-mer cages using substrates with varying sequences and lengths [10], [11], [13], [14], in our case, we first tested our hypothesis using substrates that only vary in length.
With the sulfinate radical chemistry, we were confident that we would be able to tag traditionally "untaggable" substrates that only contain C H bonds.
Similar(58)
Although chitinases and chitosanases hydrolyze chemically similar substrates that differ only by an acetyl group, no sequence similarities were found between members of these two families.
Two DNA substrates that differed only in the base to the 5′ side of the adduct were used (Table 1, substrates 1 and 2).
To test the impact of the central four base pair sequence, we compared cleavage of DNA substrates that differ only at these center four base pairs.
Based on the polarity of the DNA at the active site, these residues are predicted to contact regions of flap and splayed arm substrates that contain only double-stranded DNA.
Indeed, the proteasome has clear amino acid sequence preferences as degradation of model substrates that differ only in their disordered initiation regions varies over at least an order of magnitude (Fishbain et al., 2014).
Synthetic substrates that differ only in leader (for example benzyloxycarbonyl, succinyl or tosyl) or reporter groups (for example aminomethylcoumarin, naphthylamide or nitroanilide) are considered different cleavages even if all are performed by the same peptidase.
ACE activity was tested, both in cell lysates and in cell culture medium, using substrates that were only moderately domain selective, i.e. HHL (Hip-His-Leu) and AcSDAcKP (Acetyl-Serine-Aspartyl-Acetyl-Lysine-Proline), a C- and an N-domain substrate, respectively.
To our knowledge, GST-Kre22 is the first substrate that requires only Dsk2, but not other Ub receptors (e.g. Rpn10, Rad23) for degradation.
To our knowledge, Kre22 is the first substrate that requires only Dsk2 but none of the other Ub receptors for its degradation.
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