Abstract
BackgroundPeptide aptamers are combinatorial protein reagents that bind to targets with a high specificity and a strong affinity thus providing a molecular tool kit for modulating the function of their targets in vivo.ResultsHere we report the isolation of a peptide aptamer named swiggle that interacts with the very short (21 amino acid long) intracellular domain of membrane type 1-metalloproteinase (MT1-MMP), a key cell surface protease involved in numerous and crucial physiological and pathological cellular events. Expression of swiggle in mammalian cells was found to increase the cell surface expression of MT1-MMP by impairing its internalisation. Swiggle interacts with the LLY573 internalisation motif of MT1-MMP intracellular domain, thus disrupting the interaction with the μ2 subunit of the AP-2 internalisation complex required for endocytosis of the protease. Interestingly, swiggle-mediated inhibition of MT1-MMP clathrin-mediated internalisation was also found to promote MT1-MMP-mediated cell migration.ConclusionsTaken together, our results provide further evidence that peptide aptamers can be used to dissect molecular events mediated by individual protein domains, in contrast to the pleiotropic effects of RNA interference techniques.
Highlights
Peptide aptamers are combinatorial protein reagents that bind to targets with a high specificity and a strong affinity providing a molecular tool kit for modulating the function of their targets in vivo
Isolation of peptide aptamers that interact with the membrane-type I matrix metalloproteinase (MT1-MMP) intracellular domain (ICD) In order to identify PAs that interact with the MT1MMP ICD, a yeast two-hybrid screen was performed as previously described [1]
The 21 amino acid ICD of MT1-MMP was fused to the LexA DNA-binding domain (DBD) to generate the bait (LexA-MT1)
Summary
Peptide aptamers are combinatorial protein reagents that bind to targets with a high specificity and a strong affinity providing a molecular tool kit for modulating the function of their targets in vivo. Peptide aptamers (PAs) are small, artificially engineered proteins conceptually similar to antibodies [1]. PAs consist of a stable, ideally inert scaffold protein with an inserted constrained peptide moiety. This in effect presents a small peptide surface within the tertiary structure of the scaffold which serves as the binding site for a target protein. Some PAs have been found to block functions of their target proteins in vivo, such as human Cdk2 [10], D. melanogaster Cdk and 2 [3], E2F [11], p53 [12], Stat3 [8], Nr-13 [13], and BCL-6 [9]
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