title: Effect of ELP Sequence and Fusion Protein Design on Concentrated Solution Self-Assembly.
abstract: Fusion proteins provide a facile route for the purification and self-assembly of biofunctional protein block copolymers into complex nanostructures; however, the use of biochemical synthesis techniques introduces unexplored variables into the design of the structures. Using model fusion constructs of the red fluorescent protein mCherry and the coil-like protein elastin-like polypeptide (ELP), it is shown that the molar mass and hydrophobicity of the ELP sequence have a large effect on the propensity of a fusion to form well-ordered nanostructures, even when the ELP is in the low temperature, highly solvated state. In contrast, the presence of a 6xHis purification tag has little effect on self-assembly, and the order of blocks in the construct (N-terminal vs C-terminal) only has a significant effect on the nanostructure when the conjugates are heated above the transition temperature of the ELP block. These results indicate that for a sufficiently hydrophobic and high molar mass ELP block, there is a great deal of design latitude in the construction of fusion protein block copolymers for self-assembling nanomaterials.
authors.affiliation: Department of Chemical Engineering, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.
authors.forename: Guokui
authors.initials: G
authors.lastname: Qin
authors.affiliation: Department of Chemical Engineering, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.
authors.forename: Paola M
authors.initials: PM
authors.lastname: Perez
authors.affiliation: Department of Chemical Engineering, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.
authors.forename: Carolyn E
authors.initials: CE
authors.lastname: Mills
authors.affiliation: Department of Chemical Engineering, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.
authors.forename: Bradley D
authors.initials: BD
authors.lastname: Olsen
mesh_terms: D057927:Hydrophobic and Hydrophilic Interactions; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics; D049329:Nanostructures / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry* / Q000235:genetics; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D000097573:Red Fluorescent Protein
chemical_list: D008164:Luminescent Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein; C033943:elastin polypentapeptide