Symmetry Controlled, Genetic Presentation of Bioactive Proteins on the P22 Virus-like Particle Using an External Decoration Protein.
Viruses use spatial control of constituent proteins as a means of manipulating and evading host immune systems. Similarly, precise spatial control of proteins encapsulated or presented on designed nanoparticles has the potential to biomimetically amplify or shield biological interactions. Previously, we have shown the ability to encapsulate a wide range of guest proteins within the virus-like particle (VLP) from Salmonella typhimurium bacteriophage P22, including antigenic proteins from human pathogens such as influenza. Expanding on this robust encapsulation strategy, we have used the trimeric decoration protein (Dec) from bacteriophage L as a means of controlled exterior presentation on the mature P22 VLP, to which it binds with high affinity. Through genetic fusion to the C-terminus of the Dec protein, either the 17 kDa soluble region of murine CD40L or a minimal peptide designed from the binding region of the "self-marker" CD47 was independently presented on the P22 VLP capsid exterior. Both candidates retained function when presented as a Dec-fusion. Binding of the Dec domain to the P22 capsid was minimally changed across designed constructs, as measured by surface plasmon resonance, demonstrating the broad utility of this presentation strategy. Dec-mediated presentation offers a robust, modular means of decorating the exposed exterior of the P22 capsid in order to further orchestrate responses to internally functionalized VLPs within biological systems.
Department of Chemistry, Indiana University , 800 E. Kirkwood Avenue, Bloomington, Indiana 47405, United States.
Benjamin
B
Schwarz
Department of Chemistry and Biochemistry, Montana State University , PO Box 173400, Bozeman, Montana 59717, United States.
Patrick
P
Madden
Department of Chemistry, Indiana University , 800 E. Kirkwood Avenue, Bloomington, Indiana 47405, United States.
John
J
Avera
BiOptix Inc. , 1775 38th Street, Boulder, Colorado 80301, United States.
Bridget
B
Gordon
Department of Microbiology and Immunology, Montana State University , PO Box 173520, Bozeman, Montana 59717, United States.
Kyle
K
Larson
Department of Microbiology and Immunology, Montana State University , PO Box 173520, Bozeman, Montana 59717, United States.
Heini M
HM
Miettinen
Department of Chemistry, Indiana University , 800 E. Kirkwood Avenue, Bloomington, Indiana 47405, United States.
Masaki
M
Uchida
Department of Chemistry and Biochemistry, Montana State University , PO Box 173400, Bozeman, Montana 59717, United States.
Ben
B
LaFrance
Department of Biophysics, Bose Institute , Kolkata 700054, India.
Gautam
G
Basu
Department of Microbiology and Immunology, Montana State University , PO Box 173520, Bozeman, Montana 59717, United States.
Agnieszka
A
Rynda-Apple
Department of Chemistry, Indiana University , 800 E. Kirkwood Avenue, Bloomington, Indiana 47405, United States.
Trevor
T
Douglas
D000818:Animals; D017100:Bacteriophage P22 / Q000737:chemistry* / Q000235:genetics; D010582:Bacteriophage lambda / Q000737:chemistry / Q000235:genetics; D023201:CD40 Ligand / Q000737:chemistry / Q000235:genetics; D051928:CD47 Antigen / Q000737:chemistry / Q000235:genetics; D036022:Capsid Proteins / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D051379:Mice; D010455:Peptides / Q000737:chemistry / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics*; D012486:Salmonella typhimurium / Q000821:virology; D014771:Virion / Q000737:chemistry* / Q000235:genetics
D051928:CD47 Antigen; D036022:Capsid Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; D023201:CD40 Ligand
CD40L (CD154); SPR; bacteriophage L decoration protein (dec); bacteriophage P22; polyvalent display; virus-like particle