The DYKDDDDK-tag system utilizes a short, hydrophilic 8- amino-acid peptide that is fused to the protein of interest1. The DYKDDDDK peptide binds to the antibody M1. Whether binding is calcium-dependent manner2 or –independent3 remains controversial. A disadvantage of the system is that the monoclonal antibody purification matrix is not as stable as others. In general, small tags can be detected with specific monoclonal antibodies.
To improve the detection of the DYKDDDDK tag the 3x (DYKDDDDK) system has been developed. This three tandem DYKDDDDK epitope is hydrophilic, 22-amino-acids long and can detect up to 10 fmol of expressed fusion protein. The DYKDDDDK-tagged maltodextrin-binding protein of Pyrococcus furiosus has been crystallized4 and the quality of the crystals was very similar to that of crystals of untagged protein.
Finally, the DYKDDDDK-tag can be removed by treatment with enterokinase, which is specific for the five C-terminal amino acids of the peptide sequence5.
References:
1. Hopp TP, Prickett KS, Price VL, Libby RT, March CJ,
Ceretti DP, Urdal DL, Conlon PJ (1988) A short polypeptide marker
sequence useful for recombinant protein identification and purification.
Bio/Technology 6:1204–1210.
2. Hopp TP, Gallis B, Prikett KS (1996)
Metal-binding properties of a calcium dependent monoclonal antibody. Mol
Immunol 33:601–608.
3. Einhauer A, Jungbauer A (2000) Kinetics and
thermodynamical properties of the monoclonal antibody M1 directed
against the FLAG peptide. 20th International symposium on the separation
of proteins, peptides, and polynucleotides (ISPPP). Lublijana,
Slovenia, November 5–8, 2000.
4. Bucher MH, Evdokimov AG, Waugh DS
(2002) Differential effects of short affinity tags on the
crystallization of Pyrococcus furiosus maltodextrin-binding protein.
Biol Cryst 58:392–397.
5. Maroux S, Baratti J, Desnuelle P (1971) Purification and specificity of procine enterokinase. J Biol Chem 246:5031–5039.
Product Citation
- 1. Yao Gong, Gaurav Behera, et al. "HypDB: A functionally annotated web-based database of the proline hydroxylation proteome." PLoS Biol. 2022 Aug 26;20(8):e3001757. PMID: 36026437
- 2. Daniel Frank, Sarah E Garnish, et al. "Ubiquitylation of RIPK3 beyond-the-RHIM can limit RIPK3 activity and cell death." iScience. 2022 Jun 17;25(7):104632. PMID: 35800780
- 3. Md Mahfuz Al Mamun, Muhammad Riaz Khan, et al. "Stub1 maintains proteostasis of master transcription factors in embryonic stem cells." Cell Rep. 2022 Jun 7;39(10):110919. PMID: 35675767
- 4. Yang Xu, Han Han, et al. "Active conformation of the p97-p47 unfoldase complex." Nat Commun. 2022 May 12;13(1):2640. PMID: 35552390
- 5. Lin Liu, Jarrod J. Sandow, et al. "Tankyrase-mediated ADP-ribosylation is a regulator of TNF-induced death." Sci Adv. 2022 May 13;8(19):eabh2332. PMID: 35544574
- 6. Michael G Hanna, Patreece H Suen, et al. "SHIP164 is a chorein motif lipid transfer protein that controls endosome–Golgi membrane traffic." J Cell Biol. 2022 Jun 6;221(6):e202111018. PMID: 35499567
Chemical Properties
| Storage | Desiccate at -20°C |
| M.Wt | 2861.87 |
| Cas No. | 402750-12-3 |
| Formula | C120H169N31O49S |
| Synonyms | H-Met-Asp-Tyr-Lys-Asp-His-Asp-Gly-Asp-Tyr-Lys-Asp-His-Asp-Ile-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-OH |
| Solubility | ≥25mg/ml in TBS (0.5M Tris-HCl, pH 7.4, with 1M NaCl), 1X TBS as recommended buffer. |
| Chemical Name | 3X (DYKDDDDK) Peptide |
| Canonical SMILES | CCC(C)C(C(=O)NC(CC(=O)O)C(=O)NC(CC1=CC=C(C=C1)O)C(=O)NC(CCCCN)C(=O)NC(CC(=O)O)C(=O)NC(CC(=O)O)C(=O)NC(CC(=O)O)C(=O)NC(CC(=O)O)C(=O)NC(CCCCN)C(=O)O)NC(=O)C(CC(=O)O)NC(=O)C(CC2=CNC=N2)NC(=O)C(CC(=O)O)NC(=O)C(CCCCN)NC(=O)C(CC3=CC=C(C=C3)O)NC(=O)C(CC(=O)O)NC( |
| Shipping Condition | Small Molecules with Blue Ice, Modified Nucleotides with Dry Ice. |
| General tips | For obtaining a higher solubility, please warm the tube at 37°C and shake it in the ultrasonic bath for a while. We do not recommend long-term storage for the solution, please use it up soon. |