9l08

Publication Abstract from PubMed

Protein-based supramolecules require precise arrangement of building blocks. A regular-triangle trimer (cp-c(555))(3) has been constructed using an alpha-helix-inserted-circular permutant (cp-c(555)) of Aquifex aeolicus cytochrome (cyt) c(555), where the trimers may dissociate to monomers. In this study, we stabilized the regular-triangle structure by constructing a cyclic regular-triangle of three alpha-helix-linked cyt c(555) molecules using sortase-mediated ligation (SML). Comparing SML using sortase A for six cp-c(555) variant trimers, the variant with GGG at the N-terminus and LPETG at the C-terminus reacted most efficiently. OP-(c(555))(3) was designed, in which two cyt c(555) molecules were fused using an alpha-helix, generating a dimer. The cyt c(555) C-terminal region was attached to the N-terminus of the dimer, and the cyt c(555) N-terminal region was attached to the C-terminus of the dimer using the same alpha-helix. OP-(c(555))(3) was expressed in Escherichia coli cells, and the termini were connected by SML, forming a cyclic regular-triangle, CL-(c(555))(3). CL-(c(555))(3) showed higher thermostability than (cp-c(555))(3) and OP-(c(555))(3). CL-(c(555))(3) structural stability was confirmed using high-speed atomic force microscopy. The X-ray crystal structure of CL-(c(555))(3) showed a cyclic structure and a nanoporous supramolecular assembly. These results demonstrate that a nanoporous supramolecular assembly can be constructed by designing a cyclic molecule with a central hole using SML.

Construction of a Cyclic Regular-Triangle Trimer of Cytochrome c(555) with a Central Hole Using Sortase A.,Novientri G, Fujiwara K, Mashima T, Matsuura H, Ogata H, Uchihashi T, Fujii S, Sambongi Y, Hirota S Chemistry. 2025 May 14;31(27):e202404736. doi: 10.1002/chem.202404736. Epub 2025 , Apr 16. PMID:40148242^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.