Quartz 4

Heterodimerization domains

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Heterodimerization domains can be used to noncovalently link two different proteins together.

Examples

  • Barnase/barstar (1) is a high-affinity protein pair used for exactly these purposes. That review puts the affinity (Kd) at the femtomolar level. Someone suggested introducing a mutation in the active site of barnase, which is an active ribonuclease; Mossakowska et al. (2) suggest H102A and E73A, but I’m unclear on whether that also prevents barstar binding. More research is needed.
  • Colicin E9DNAse/Im9 Cognate Immunity Protein, which is also femtomolar in binding affinity (3). Again, Colicin is an active enzyme, so some search for mutations may be necessary.
  • Trypsin/BPTI and anhydrotrypsin/BPTI, also femtomolar (4).
  • Choice mutations in OMTKY3/SGBP allow the binding affinity to allegedly reach subfemtomolar affinity (5).
  • SpyCatcher/SpyTag (6).
  • FKBP (12 kDa FK506 binding protein) and FRB, which is rapamycin-dependent (citation needed)
  • Fc-regions
1.
Shilova O, Kotelnikova P, Proshkina G, Shramova E, Deyev S. Barnase-Barstar Pair: Contemporary Application in Cancer Research and Nanotechnology. Molecules. 2021;26(22):6785. Available from: https://doi.org/10.3390/molecules26226785
2.
Mossakowska DE, Nyberg K, Fersht AR. Kinetic characterization of the recombinant ribonuclease from Bacillus amyloliquefaciens (barnase) and investigation of key residues in catalysis by site-directed mutagenesis. Biochemistry. 1989;28(9):3843–50. Available from: https://doi.org/10.1021/bi00435a033
3.
Wallis R, Leung K-Y, Pommer AJ, Videler H, Moore GR, James R, et al. Protein-Protein Interactions in Colicin E9 DNase-Immunity Protein Complexes. 2. Cognate and Noncognate Interactions That Span the Millilmolar to Femptomolar Affinity Range. Biochemistry. 1995;34(42):13751–9. Available from: https://doi.org/10.1021/bi00042a005
4.
Borjigin J, Nathans J. Bovine pancreatic trypsin inhibitor-trypsin complex as a detection system for recombinant proteins. Proceedings of the National Academy of Sciences. 1993;90(1):337–41. Available from: https://doi.org/10.1073/pnas.90.1.337
5.
Qasim MA, Wang L, Qasim S, Lu S, Lu W, Wynn R, et al. Additivity‐based design of the strongest possible turkey ovomucoid third domain inhibitors for porcine pancreatic elastase (PPE) andStreptomyces griseusprotease B (SGPB). FEBS Letters. 2013;587(18):3021–6. Available from: https://doi.org/10.1016/j.febslet.2013.07.029
6.
Fierle JK, Abram-Saliba J, Brioschi M, deTiani M, Coukos G, Dunn SM. Integrating SpyCatcher/SpyTag covalent fusion technology into phage display workflows for rapid antibody discovery. Scientific Reports. 2019;9(1). Available from: https://doi.org/10.1038/s41598-019-49233-7

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