Activation of bioluminescence by structural complementation
View Patent ↗Provided herein are compositions and methods for the assembly of a bioluminescent complex from two or more non-luminescent (e.g., substantially non-luminescent) peptide and/or polypeptide units. In particular, bioluminescent activity is conferred upon a non-luminescent polypeptide via structural complementation with another, complementary non-luminescent peptide.
1. A peptide comprising an amino acid sequence having greater than 45% but not greater than 90% sequence identity with SEQ ID NO: 2, wherein a bioluminescent signal produced in the presence of a furimazine substrate is substantially increased when the peptide contacts a polypeptide consisting of SEQ ID NO: 440 when compared to a bioluminescent signal produced by the peptide and furimazine substrate alone, and wherein the amino acid sequence is not a naturally occurring protein or a fragment thereof.
2. The peptide of claim 1 , wherein the bioluminescent signal is substantially increased when the peptide associates with the polypeptide consisting of SEQ ID NO: 440.
3. The peptide of claim 1 , wherein the peptide exhibits enhancement of one or more traits compared to a peptide of SEQ ID NO: 2, wherein the traits are selected from: affinity for the polypeptide consisting of SEQ ID NO: 440, expression, intracellular solubility, intracellular stability, and bioluminescent activity when combined with the polypeptide consisting of SEQ ID NO: 440.
4. The peptide of claim 1 , wherein the amino acid sequence is selected from the peptides of Table 1.
5. The peptide of claim 1 , wherein the amino acid sequence is synthetic, contains non-natural amino acids or is a peptide mimic.
6. A fusion polypeptide comprising the peptide of claim 1 and a first interaction polypeptide that is configured to form a complex with a second interaction polypeptide upon contact of the first interaction polypeptide and the second interaction polypeptide.
7. A bioluminescent complex comprising:
(a) the fusion polypeptide of claim 6 ; and
(b) a second fusion polypeptide comprising:
(i) the second interaction polypeptide, and
(ii) a complement polypeptide that emits a detectable bioluminescent signal in the presence of a substrate when associated with a peptide comprising an amino acid sequence having less than 100% and greater than 40% sequence identity with SEQ ID NO: 2;
wherein the fusion polypeptide of claim 6 and the second fusion polypeptide are associated;
and wherein the peptide comprising an amino acid sequence having greater than 45% but not greater than 90% sequence identity with SEQ ID NO: 2 and the complement polypeptide are associated.
8. A bioluminescent complex comprising a non-covalent association of:
(a) a peptide comprising a peptide amino acid sequence having greater than 45% but not greater than 90% sequence identity with SEQ ID NO: 2, wherein the peptide amino acid sequence is not a naturally occurring protein or a fragment thereof; and
(b) a polypeptide comprising a polypeptide amino acid sequence having greater than 40% but not greater than 95% sequence identity with SEQ ID NO: 440, wherein the polypeptide amino acid sequence is not a naturally occurring protein or a fragment thereof, and wherein the bioluminescent complex exhibits detectable luminescence;
wherein the bioluminescent complex produces substantially increased luminescence in the presence of a furimazine substrate when compared to either the peptide or polypeptide alone in the presence of the furimazine substrate.
9. The bioluminescent complex of claim 8 , wherein the peptide amino acid sequence is selected from the peptide sequences of Table 1.
10. The bioluminescent complex of claim 8 , wherein the polypeptide amino acid sequence is selected from the polypeptide sequences of Table 2.
11. The bioluminescent complex of claim 8 , wherein the peptide amino acid sequence is attached to a first interaction element that is associated with a second interaction element.
12. The bioluminescent complex of claim 11 , wherein the polypeptide amino acid sequence is attached to the second interaction element.
13. The bioluminescent complex of claim 12 , wherein the polypeptide amino acid sequence and peptide amino acid sequence are substantially non-luminescent in the absence of the association of the first and second interaction elements.