6I2G

ALFA-tag binding nanobody (NbALFA) bound to ALFA-tag peptide.

Summary for 6I2G

• Entry DOI: 10.2210/pdb6i2g/pdb
• Descriptor: ALFA nanobody, N7P-SER-ARG-LEU-GLU-GLU-GLU-LEU-ARG-ARG-ARG-LEU-THR-GLU-LPD, SULFATE ION, ... (4 entities in total)
• Functional Keywords: nanobody, alfa, tag, peptide binding protein
• Biological source: Vicugna pacos; More
• Total number of polymer chains: 2
• Total formula weight: 15717.49

Authors

Martinez-Carranza, M.,Stenmark, P. (deposition date: 2018-11-01, release date: 2019-09-25, Last modification date: 2024-01-24)

Primary citation

Gotzke, H.,Kilisch, M.,Martinez-Carranza, M.,Sograte-Idrissi, S.,Rajavel, A.,Schlichthaerle, T.,Engels, N.,Jungmann, R.,Stenmark, P.,Opazo, F.,Frey, S.
The ALFA-tag is a highly versatile tool for nanobody-based bioscience applications.
Nat Commun, 10:4403-4403, 2019

PubMed Abstract: Specialized epitope tags are widely used for detecting, manipulating or purifying proteins, but often their versatility is limited. Here, we introduce the ALFA-tag, a rationally designed epitope tag that serves a remarkably broad spectrum of applications in life sciences while outperforming established tags like the HA-, FLAG®- or myc-tag. The ALFA-tag forms a small and stable α-helix that is functional irrespective of its position on the target protein in prokaryotic and eukaryotic hosts. We characterize a nanobody (NbALFA) binding ALFA-tagged proteins from native or fixed specimen with low picomolar affinity. It is ideally suited for super-resolution microscopy, immunoprecipitations and Western blotting, and also allows in vivo detection of proteins. We show the crystal structure of the complex that enabled us to design a nanobody mutant (NbALFA) that permits efficient one-step purifications of native ALFA-tagged proteins, complexes and even entire living cells using peptide elution under physiological conditions.

PubMed: 31562305
DOI: 10.1038/s41467-019-12301-7

Experimental method

X-RAY DIFFRACTION (1.5 Å)