Part:BBa_K5466002

AGA2P Signal Peptide

Signal peptide consisting on the 18 first amino acids from Saccharomyces cerevisiae AGA2 protein. Attachment to the N-terminal end of a protein can efficiently drive translocation across the plasma membrane. Additional fusion of AGA2P to the C-terminal can be used for protein display.

AG was added to the C-terminus of the signal peptide based on the discovery by Wang et al. (2005) regarding the importance of specific amino acids for proper peptide cleavage. More details can be found on the design page.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

AGA2P

AGA2 is an extracellularly-secreted glycoprotein that constitutes the adhesion subunit of a-agglutinin in a-cells. The N-terminal sequence of the protein harbors a signal peptide (SP) required for protein translocation. Its C-terminal sequence acts as a ligand for alpha-agglutinin, promoting binding and aggregation during mating.

AGA2 remains strongly anchored to the cell wall thanks to AGA1, the anchorage subunit of a-agglutinin. AGA1 stays attached to the cell surface through GPI and strongly binds AGA2 via two disulfide bonds.

Proten display

Previously applied display strategies involved fusion of AGA2P and its signal peptide to the N-terminus of the protein of interest. Wang et al. (2005) showed increased affinity constants for displayed scFvs when AGA2 signal peptide was attached to the N-terminal and the rest of the protein was fused to the C-terminal end of the scFv.

Reference

Wang, Z., Mathias, A., Stavrou, S., & Neville, D. M. (2005). A new yeast display vector permitting free scFv amino termini can augment ligand binding affinities. Protein Engineering Design And Selection, 18(7), 337-343. https://doi.org/10.1093/protein/gzi036

Zhao, H., Shen, Z. M., Kahn, P. C., & Lipke, P. N. (2001). Interaction of alpha-agglutinin and a-agglutinin, Saccharomyces cerevisiae sexual cell adhesion molecules. Journal of bacteriology, 183(9), 2874–2880. https://doi.org/10.1128/JB.183.9.2874-2880.2001