TY - JOUR
T1 - Engineered domain swapping as an on/off switch for protein function
AU - Ha, Jeung Hoi
AU - Karchin, Joshua M.
AU - Walker-Kopp, Nancy
AU - Castañeda, Carlos A.
AU - Loh, Stewart N.
N1 - Publisher Copyright:
© 2015 Elsevier Ltd All rights reserved.
PY - 2015/10/22
Y1 - 2015/10/22
N2 - Domain swapping occurs when identical proteins exchange segments in reciprocal fashion. Natural swapping mechanisms remain poorly understood, and engineered swapping has the potential for creating self-assembling biomaterials that encode for emergent functions. We demonstrate that induced swapping can be used to regulate the function of a target protein. Swapping is triggered by inserting a "lever" protein (ubiquitin) into one of four loops of the ribose binding protein (RBP) target. The lever splits the target, forcing RBP to refold in trans to generate swapped oligomers. Identical RBP-ubiquitin fusions form homo-swapped complexes with the ubiquitin domain acting as the hinge. Surprisingly, some pairs of non-identical fusions swap more efficiently with each other than they do with themselves. Nuclear magnetic resonance experiments reveal that the hinge of these hetero-swapped complexes maps to a region of RBP distant from both ubiquitins. This design is expected to be applicable to other proteins to convert them into functional switches.
AB - Domain swapping occurs when identical proteins exchange segments in reciprocal fashion. Natural swapping mechanisms remain poorly understood, and engineered swapping has the potential for creating self-assembling biomaterials that encode for emergent functions. We demonstrate that induced swapping can be used to regulate the function of a target protein. Swapping is triggered by inserting a "lever" protein (ubiquitin) into one of four loops of the ribose binding protein (RBP) target. The lever splits the target, forcing RBP to refold in trans to generate swapped oligomers. Identical RBP-ubiquitin fusions form homo-swapped complexes with the ubiquitin domain acting as the hinge. Surprisingly, some pairs of non-identical fusions swap more efficiently with each other than they do with themselves. Nuclear magnetic resonance experiments reveal that the hinge of these hetero-swapped complexes maps to a region of RBP distant from both ubiquitins. This design is expected to be applicable to other proteins to convert them into functional switches.
UR - http://www.scopus.com/inward/record.url?scp=84950302300&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84950302300&partnerID=8YFLogxK
U2 - 10.1016/j.chembiol.2015.09.007
DO - 10.1016/j.chembiol.2015.09.007
M3 - Article
C2 - 26496687
AN - SCOPUS:84950302300
SN - 1074-5521
VL - 22
SP - 1384
EP - 1393
JO - Chemistry and Biology
JF - Chemistry and Biology
IS - 10
ER -