Molecular building blocks are fundamental to biological synthesis and processes and have been utilized in advanced materials, drugs and drug delivery systems, and biotechnology. Proteins have been used as molecular building blocks for the construction of complex, well-ordered structures. Coiled-coil protein domains are essential subunits used for the oligomerization of protein complexes, gene expression, and structural elements of biological materials. The synthesis and assembly of proteins utilizing coiled-coil motifs are of great scientific interest due to their potential applications in disease treatment, biomechanical motors, nanoscale delivery systems, etc. However, assembling protein complexes with specific morphology is still challenging because the controllability of the protein association is complicated by multiple interactions between a diverse array of amino acids.
Initial progress toward the design, synthesis, and characterization of paired coiled-coil peptidic molecular building blocks that can self-assemble with a high degree of controllability are presented herein. Eight unique 32-residue peptides were synthesized, each with one modified residue to covalently crosslink two peptides via a 1,3-dipolar cycloaddition click reaction. Each peptide was expected to fold into α-helical coiled-coil heterodimer peptide pairs upon association with its complementary pair. Furthermore, each crosslinked MBB was designed to exhibit controlled self-assembly with its specific complementary MBB. This design strategy allowed the intermolecular interactions of the peptide to control self-assembly of multiple MBBs via association through the sidechains of the heptadic sequences. Mass spectroscopy, size-exclusion chromatography (SEC), and HPLC analysis show successful synthesis and purification of individual peptides. Additional chromatographic data show successful association of dimeric coiled coils, synthesis of crosslinked MBBs, and self-assembly of the crosslinked MBBs.
Date of publication
Sean C. Butler, Dustin P. Patterson, Rachel N. Mason
Master of Science in Chemistry
DiStefano, Jason, "DESIGN, SYNTHESIS, AND ANALYSIS OF PAIRED COILED-COIL PEPTIDIC MOLECULAR BUILDING BLOCKS USED FOR LINEARLY CONTROLLED SELF-ASSEMBLY OF α-HELICAL COILED-COIL HETERODIMER PEPTIDE PAIRS" (2022). Chemistry Theses. Paper 2.