In molecular biology, alanine scanning is a site-directed mutagenesis technique used to determine the contribution of a specific residue to the stability or function of a given protein. [1] Alanine is used because of its non-bulky, chemically inert, methyl functional group that nevertheless mimics the secondary structure preferences that many ...

2021年4月20日 · Learn about how alanine scanning mutagenesis helps to map amino acid residues essential for protein functions and protein-protein interactions facilitating immunogen and antibody design and supporting therapeutic development.

Single amino acid scanning mutagenesis is a widely used method for identifying protein positions that are important for function or ligand binding. Alanine is often employed, and was selected on rational grounds as it constitutes a deletion of the side chain at the β-carbon.

2021年4月21日 · Among different approaches, alanine scanning mutagenesis is a versatile strategy that allows researchers to evaluate the role and relevance of specific amino acid sidechains within protein regions of interest (Weiss et al. 2000).

2001年6月1日 · Alanine-scanning mutagenesis, a method of systematic alanine substitution, has been particularly useful for the identification of functional epitopes. Substitution with alanine removes all sidechain atoms past the β-carbon.

Alanine scanning mutagenesis has been used in cases of human growth hormone (hGH) binding to hGH-binding protein (hGHbp), CD4 binding to HIV-gp 120, the enzymatic activity of kinases and lysozyme stability.

Overall, this protocol enabled us to efficiently produce several alanine-scanning libraries of 400 single-point mutations with complete coverage of the protein sequence. Keywords: Scanning mutagenesis, Site-directed, High throughput mutagenesis, Two-fragment PCR, Gibson assembly.

1997年1月1日 · Our alanine stretch mutagenesis method can be used as a probe to rapidly scan a whole protein sequence in search of secondary structures or to characterize the catalytic or functional role of a stretch of residues. It also provides a …

2017年7月28日 · Here we present a high-throughput mutagenesis pipeline which allowed us to systematically generate alanine scanning libraries of 400 single-point mutations in 6 weeks, with a complete coverage...

2020年1月13日 · Alanine-scanning mutagenesis is an experimental technique where residues in a protein of interest are sequentially mutated to alanine (Cunningham and Wells, 1989). The aim is to identify those residues that contribute the most to the free energy of interaction.