Structure analysis of high-molecular weight intrinsically disordered proteins using 13C detection NMR approach

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We analyzed one of the general transcription factors in yeast cell, Tfa2, with the 13C and 1H detection NMR methods. In this work, we particularly focused on the intrinsically disordered part in Tfa2, which binds to Gal11 subunit in the Mediator complex. The severe signal overlaps typically found for the ID segment were solved using the 13C and 15N evolution axes, whose chemical shifts have significant dispersions over the other nuclear spins in protein.We found the Mediator binding domain of Tfa2, Tfa2-mbd, transiently forms three-helix structure. The transient structure formation was mediated by its homo-dimer formation. We also found that the transiently folded structure has binding ability to Gal11; the mutant that lacks the transient folding ability does not bind to Gal11. Over all, the present research has demonstrated the unique structural and functional properties associated with the ID part in Tfa2.