Difference between revisions of "Part:BBa J176131"

Revision as of 08:24, 21 October 2021

PLrigid

(EAAAR)4 peptide linker, rigid; alpha helix secondary structure

Usage and Biology

PLrigid is a 20 a.a. peptide that is based on a previously characterized alpha-helix motif (EAAAR) (Merutka et al., 1991; Sommese et al., 2010). This BioBrick part is based on work reported by Yan et al., 2007, where they used this motif to build transcription factors with tandem DNA binding domains spaced apart by rigid linkers. Alpha helices are thought to be rigid structures that restrict the position of one end relative to the other. Lack of "bendability" keeps the two proteins attached at either end from coming closer together. Since the proteins are also unable to move farther apart when they are attached with a rigid linker, the relative position of the two proteins is fixed in space.

This linker was used in a study related to small molecule-mediated protein-protein interactions, where it is important to understand the abnormal signal transduction pathways in a variety of disorders, as well as to optimize the process of drug development and validation. The paper cites that the highest signal to background ratio was achieved for the split luciferase complementation system - Renilla based on fusion protein when linker peptides containing amino acids from EAAAR repeats were chosen. It was found that peptide linker with two or four EAAAR repeats showed greater signal mediated by protein-protein interaction with lower background signal compared to no linker or linker with amino acid sequences GGGGSGGGGS, ACGSLSCGSF, and ACGSLSCGS-FACGSLSCGSF. This linker was also cited in the US patent application for CAS9 FUSION MOLECULES, GENE EDITING SYSTEMS, AND METHODS OF USING THEM (Request # 20190136210) as an additional linker.

Isoelectric Point: 6.39

Molecular Weight: 2012.17Da

REFERENCES

1. Merutka G, Shalongo W, Stellwagen E. (1991) A model peptide with enhanced helicity. Biochem. 30: 4245-4248.
2. Sommese RF, Sivaramakrishnan S, Baldwin RL, Spudich JA. (2010) Helicity of short E-R/K peptides. Protein Sci. 19: 2001-2005.
3. Yan W, Imanishi M, Futaki S, Sugiura Y. (2007) alpha-Helical Linker of an Artificial 6-Zinc Finger Peptide Contributes to Selective DNA Binding to a Discontinuous Recognition Sequence. Biochem. 46: 8517-8524.

Sequence and Features