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Sequence requirements for the recognition of tyrosine-based endocytic signals by clathrin AP-2 complexes.

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  • 1. Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
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    • Boll W 1
    (1 author)

The EMBO Journal, 01 Nov 1996, 15(21):5789-5795
https://doi.org/10.1002/j.1460-2075.1996.tb00965.x PMID: 8918456 PMCID: PMC452326

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Abstract 

We recently determined that fusion proteins containing tyrosine-based endocytic signals bind to the mu 2 subunit of AP-2, the complex that drives clathrin coat formation and mediates endocytosis from the plasma membrane. Here we analyze the selectivity of peptide recognition by mu 2 and by AP-2 using combinatorial selection methods and surface plasmon resonance. Both mu 2 and AP-2 are shown to interact with various sequences of the form tyrosine-polar-polar-hydrophobic (Yppø) found on receptors that follow the clathrin pathway. The optimal sequence for interaction with mu 2 and with AP-2 has tyrosine as an anchor and prefers arginine at position Y + 2 and leucine at position Y + 3. In contrast, no preferred sequence is detected surrounding the Yppø signal, indicating that recognition of the Yppø endocytic signal does not require a prefolded structure. We conclude that sorting into the endocytic pathway is governed by a surprisingly simple interaction between the mu 2 chain and a tyrosine-containing tetrapeptide sequence.

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EMBO J. 1996 Nov 1; 15(21): 5789–5795.
PMCID: PMC452326
PMID: 8918456

Sequence requirements for the recognition of tyrosine-based endocytic signals by clathrin AP-2 complexes.

W Boll, H Ohno, Z Songyang, I Rapoport, L C Cantley, J S Bonifacino, and T Kirchhausen
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Abstract

We recently determined that fusion proteins containing tyrosine-based endocytic signals bind to the mu 2 subunit of AP-2, the complex that drives clathrin coat formation and mediates endocytosis from the plasma membrane. Here we analyze the selectivity of peptide recognition by mu 2 and by AP-2 using combinatorial selection methods and surface plasmon resonance. Both mu 2 and AP-2 are shown to interact with various sequences of the form tyrosine-polar-polar-hydrophobic (Yppø) found on receptors that follow the clathrin pathway. The optimal sequence for interaction with mu 2 and with AP-2 has tyrosine as an anchor and prefers arginine at position Y + 2 and leucine at position Y + 3. In contrast, no preferred sequence is detected surrounding the Yppø signal, indicating that recognition of the Yppø endocytic signal does not require a prefolded structure. We conclude that sorting into the endocytic pathway is governed by a surprisingly simple interaction between the mu 2 chain and a tyrosine-containing tetrapeptide sequence.

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Selected References

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