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  • 1.
    Jespersen, Nathan
    et al.
    Oregon State Univ, Dept Biochem & Biophys, Corvallis, OR 97331 USA.
    Estelle, Aidan
    Oregon State Univ, Dept Biochem & Biophys, Corvallis, OR 97331 USA.
    Waugh, Nathan
    Oregon State Univ, Dept Biochem & Biophys, Corvallis, OR 97331 USA.
    Davey, Norman E.
    Univ Coll Dublin, Conway Inst Biomol & Biomed Sci, Dublin, Ireland.
    Blikstad, Cecilia
    Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - BMC.
    Ammon, York-Christoph
    Univ Utrecht, Dept Biol, Utrecht, Netherlands.
    Akhmanova, Anna
    Univ Utrecht, Dept Biol, Utrecht, Netherlands.
    Ivarsson, Ylva
    Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - BMC, Biochemistry.
    Hendrix, David A.
    Oregon State Univ, Dept Biochem & Biophys, Corvallis, OR 97331 USA;Oregon State Univ, Sch Elect Engn & Comp Sci, Corvallis, OR 97331 USA.
    Barbar, Elisar
    Oregon State Univ, Dept Biochem & Biophys, Corvallis, OR 97331 USA.
    Systematic identification of recognition motifs for the hub protein LC82019In: LIFE SCIENCE ALLIANCE, ISSN 2575-1077, Vol. 2, no 4, article id e201900366Article in journal (Refereed)
    Abstract [en]

    Hub proteins participate in cellular regulation by dynamic binding of multiple proteins within interaction networks. The hub protein LC8 reversibly interacts with more than 100 partners through a flexible pocket at its dimer interface. To explore the diversity of the LC8 partner pool, we screened for LC8 binding partners using a proteomic phage display library composed of peptides from the human proteome, which had no bias toward a known LC8 motif. Of the identified hits, we validated binding of 29 peptides using isothermal titration calorimetry. Of the 29 peptides, 19 were entirely novel, and all had the canonical TQT motif anchor. A striking observation is that numerous peptides containing the TQT anchor do not bind LC8, indicating that residues outside of the anchor facilitate LC8 interactions. Using both LC8-binding and nonbinding peptides containing the motif anchor, we developed the "LC8Pred" algorithm that identifies critical residues flanking the anchor and parses random sequences to predict LC8-binding motifs with similar to 78% accuracy. Our findings significantly expand the scope of the LC8 hub interactome.

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