3-Azido-D-alanine HCl

Product Information

Stable and Bioorthogonal Protein Labeling with D-Amino Acid Technology

3-Azido-D-alanine hydrochloride ((R)-2-Amino-3-azidopropanoic acid hydrochloride) is a synthetic, unnatural D-amino acid derivative of alanine carrying an azide group. It enables site-specific protein modification and bioorthogonal labeling via Click Chemistry (SPAAC or CuAAC) or Staudinger ligation under mild conditions.

This reagent can be incorporated into peptides or proteins through:

• Genetic Code Expansion: Using engineered tRNAs (based on prokaryotic tRNAs) that recognize amber codons as UAG, therefore 3-Azido-D-alanine is introduced during biosynthesis at a defined position ⁽¹⁾ .
• Solid Phase Peptide Synthesis (SPPS): Allows chemical incorporation at any position in the peptide sequence.

Once incorporated, the azide moiety reacts selectively with alkyne-modified partners (alkynes, DBCO, BCN) or P(III)-containing reagents, enabling efficient conjugation without side reactions.

Why choose 3-Azido-D-alanine?

Traditional labeling methods rely on NHS ester chemistry, which reacts with primary amines (e.g., lysines) and leads to:

• Non-specific labeling at multiple sites.
• Requires basic pH (8–9) and overnight reactions.
• Risks protein degradation and structural changes.

In contrast, azide-based labeling with 3-Azido-D-alanine offers:

• Site-specific modification at the planned position.
• Fast reactions at neutral pH with high yields.
• Preserves tertiary structure.
• Fully bioorthogonal—no interference with native functional groups.
• Lower reagent consumption and atom-economic click chemistry for cost-effective, eco-friendly workflows.

Unique Benefits of D-Amino Acid Incorporation

• Increased stability: Peptides containing D-amino acids resist enzymatic degradation by proteases and peptidases.
• Structural variation: D-amino acids alter peptide folding (e.g., preventing α-helix formation), enabling novel structural designs for research and drug development.

Applications of 3-Azido-D-alanine HCl in Protein Engineering

3-Azido-D-alanine is a powerful tool for protein and peptide labeling, enabling precise site-specific modification through bioorthogonal click chemistry. It is widely used for antibody modification and ADC synthesis, supporting the development of targeted therapeutics. Researchers also rely on this reagent for linking targeting moieties to proteins in drug delivery and diagnostic applications. Additionally, its compatibility as a building block for solid-phase peptide synthesis (SPPS) makes it ideal for custom peptide design in pharmaceutical development, biotechnology, and advanced biochemistry workflows.

LITERATURE

1. Incorporating unnatural amino acids into recombinant proteins in living cells, Mitra, N., 2013, Materials and Methods, Vol. 3, p.204, 1.

http://doi.org/10.13070/mm.en.3.204

2. Mapping the Ligand-Binding Site on a G Protein-Coupled Receptor (GPCR) Using Genetically Encoded Photocrosslinkers, Grunbeck et al., 2011, Biochemistry, Vol. 50, p. 3411-3413.

https://doi.org/10.1021/bi200214r

3. Azide click chemistry on magnetotactic bacteria: A versatile technique to attach a cargo, P. E. D. S. Rodriguez et al., 2023, Materials Today Bio, Vol. 19, 100587.

https://doi.org/10.1016/j.mtbio.2023.100587

4. Novel Chromosome Organization Pattern in Actinomycetales—Overlapping Replication Cycles Combined with Diploidy, K. Böhm et al., 2017, MBio, Vol. 8(3), p. 10-1128.

https://doi.org/10.1128/mbio.00511-17

5. Azide click chemistry on magnetotactic bacteria: a versatile technique to attach a cargo, Soto, P., Sirinelli-Kojadinovic, M., Rouzaud, M., & Faivre, D., 2022, Chemrxiv.

https://doi.org/10.26434/chemrxiv-2022-dtb1t