Fig 1.
Comparison of the search process for the partner proteins that bind to the target proteins.
The well-established GST-pulldown method (left). The GST-fused bait protein is immobilized to the glutathione bead to pick up a partner protein that binds to it. The proposed aggregate pulldown (right). The ELP-fused bait protein aggregates above the transition temperature, and forms a solid support where the partner protein is immobilized by binding to the bait. By increasing the salt concentration, the partner protein is released to the solution phase while the bait-ELP still remains in the aggregate state.
Fig 2.
Disappearance and restoration of the NMR signals by the reversible aggregation.
The NMR signals originate from the partner protein, which is colored orange (left). Interaction inhibitors: competitive or non-competitive mechanisms (right). The ELP domain is not shown, but it is assumed that the ELP is aggregated into a solid phase by binding to one another. The NMR signal of the partner protein will disappear when it binds to the aggregate. The inhibitor will disrupt the binding of the partner and the target proteins. Note that the NMR spectrum in this figure is not a real data, but employed only to represent the disappearance/reappearance of the signals.
Fig 3.
The signals of partner protein is restored as it is released from the target (left). The signals of the inhibitor will disappear as it binds to the target (right). Note that the NMR spectrum in this figure is not a real data, but employed only to represent the disappearance/reappearance of the signals.