Journal: ACS nano

Article Title: Engineering the PP7 Virus Capsid as a Peptide Display Platform

doi: 10.1021/acsnano.8b09683

Figure Lengend Snippet: Sequence alignment of MS, PP7, and Qβ coat proteins. The coat protein sequences of MS2, Qβ, and PP7, aligned using the Clustal Omega35 multiple sequence alignment tool. Dashed lines mark alignment gaps, asterisks (green highlight) indicate identical residues, and single (red) and double (blue) dots indicate similar residues.

Article Snippet: The PP7 VLPs were expressed and assembled in BL21(DE3) E. coli cells (Biogen) as previously described for Q β .

Techniques: Sequencing

Journal: ACS nano

Article Title: Engineering the PP7 Virus Capsid as a Peptide Display Platform

doi: 10.1021/acsnano.8b09683

Figure Lengend Snippet: Hybrid and Homogeneous PP7 Virus-like Particles

Article Snippet: The PP7 VLPs were expressed and assembled in BL21(DE3) E. coli cells (Biogen) as previously described for Q β .

Techniques: Virus

Journal: ACS nano

Article Title: Engineering the PP7 Virus Capsid as a Peptide Display Platform

doi: 10.1021/acsnano.8b09683

Figure Lengend Snippet: Successful C-Terminal Extension Modifications of the PP7 Virus-like Particle

Article Snippet: The PP7 VLPs were expressed and assembled in BL21(DE3) E. coli cells (Biogen) as previously described for Q β .

Techniques: Virus

Journal: ACS nano

Article Title: Engineering the PP7 Virus Capsid as a Peptide Display Platform

doi: 10.1021/acsnano.8b09683

Figure Lengend Snippet: Successful Loop Insertion Modifications of the PP7-PP7 Dimer Particle

Article Snippet: The PP7 VLPs were expressed and assembled in BL21(DE3) E. coli cells (Biogen) as previously described for Q β .

Techniques: Sequencing

Journal: ACS nano

Article Title: Engineering the PP7 Virus Capsid as a Peptide Display Platform

doi: 10.1021/acsnano.8b09683

Figure Lengend Snippet: Representative analyses of thermal stability of the indicated PP7-PP7-based particles (0.1 mg/mL in PBS buffer, heated at 10 °C per minute) in the absence (top) and presence (bottom) of 1 mM DTT. Additional traces for other particles are provided in Supporting Information (Figure S1).

Article Snippet: The PP7 VLPs were expressed and assembled in BL21(DE3) E. coli cells (Biogen) as previously described for Q β .

Techniques:

Journal: ACS nano

Article Title: Engineering the PP7 Virus Capsid as a Peptide Display Platform

doi: 10.1021/acsnano.8b09683

Figure Lengend Snippet: Melting ( T m ) and Onset-of-Aggregation ( T agg ) Temperatures of PP7 Particles a

Article Snippet: The PP7 VLPs were expressed and assembled in BL21(DE3) E. coli cells (Biogen) as previously described for Q β .

Techniques:

Journal: ACS nano

Article Title: Engineering the PP7 Virus Capsid as a Peptide Display Platform

doi: 10.1021/acsnano.8b09683

Figure Lengend Snippet: Particle Sizes Measured from TEM Images

Article Snippet: The PP7 VLPs were expressed and assembled in BL21(DE3) E. coli cells (Biogen) as previously described for Q β .

Techniques:

Journal: ACS nano

Article Title: Engineering the PP7 Virus Capsid as a Peptide Display Platform

doi: 10.1021/acsnano.8b09683

Figure Lengend Snippet: Selected cryo-EM 2D class averages corresponding to the projection along the twofold axis of monomeric PP7-ZZ and ZZ-PP7 particles and dimeric PP7-PP7-ZZ particle. The apparent coronas around the first two particles are artifacts of sample preparation (scale bar = 20 nm).

Article Snippet: The PP7 VLPs were expressed and assembled in BL21(DE3) E. coli cells (Biogen) as previously described for Q β .

Techniques: Cryo-EM Sample Prep, Sample Prep

Journal: ACS nano

Article Title: Engineering the PP7 Virus Capsid as a Peptide Display Platform

doi: 10.1021/acsnano.8b09683

Figure Lengend Snippet: Cryo-EM structure of the PP7-PP7 virus-like particle [top, T = 4 (h = 2; k = 0) structure, PDB ID 6N4V] compared to the previously published structure of the PP7 particle [bottom, T = 3 (h = 1; k = 1) subunits colored the same way, PDB ID 1DWN]. (left) View down the fivefold symmetry axes; (middle) view down the twofold symmetry axis; (right) a cartoon representation showing the organization of the coat-protein dimers in the T = 4 structure. The linker loop is labeled “L”, and the termini are labeled “NC”.

Article Snippet: The PP7 VLPs were expressed and assembled in BL21(DE3) E. coli cells (Biogen) as previously described for Q β .

Techniques: Cryo-EM Sample Prep, Virus, Labeling

Journal: ACS nano

Article Title: Engineering the PP7 Virus Capsid as a Peptide Display Platform

doi: 10.1021/acsnano.8b09683

Figure Lengend Snippet: Cryo-EM structural details. (A, left) PP7-PP7 dimer threefold organization, (A, right) expanded view of one single-chain dimer, with AYGG linker (green), N-terminal Ser (cyan), and C-terminal Arg (purple) residues shown in stick representation. (B) Comparison of particles bearing the a-loop insertion and C-terminal ZZ-domain extension. Arrows mark regions of poorly resolved amino acid density in the reconstruction of the PP7-a-loop-PP7 particle following the initial Ala residue of the a-loop (left) and following the last Arg residue at the C-terminus (Arg127) (right). Circles delineate well-resolved linker loop density for the PP7-PP7-ZZ particle (left) and the Gly residue that starts the linkage to the ZZ-domain but not the rest of that domain (right).

Article Snippet: The PP7 VLPs were expressed and assembled in BL21(DE3) E. coli cells (Biogen) as previously described for Q β .

Techniques: Cryo-EM Sample Prep, Comparison, Residue

Journal: ACS nano

Article Title: Engineering the PP7 Virus Capsid as a Peptide Display Platform

doi: 10.1021/acsnano.8b09683

Figure Lengend Snippet: Mapping of extra density on the surface of PP7-a-loop-PP7, PP7-PP7-ZZ, and PP7-a-loop-PP7–150-loop particles. Overall surface density maps: (A) PP7-PP7-ZZ, (B) PP7-a-loop-PP7, and (C) PP7-a-loop-PP7–150-loop. Extra densities highlighted: (D) PP7-PP7-ZZ, (E) PP7-a-loop-PP7, and (F) PP7-a-loop-PP7–150-loop, comparing to PP7-PP7. (G) PP7-PP7-ZZ, (H) PP7-a-loop-PP7, and (I) PP7-a-loop-PP7–150-loop, showing the same loop insertions and C-terminal extensions as in (D–F). Subunit organization in (G–I) is indicated as in Figure 5.

Article Snippet: The PP7 VLPs were expressed and assembled in BL21(DE3) E. coli cells (Biogen) as previously described for Q β .

Techniques:

Journal: ACS nano

Article Title: Engineering the PP7 Virus Capsid as a Peptide Display Platform

doi: 10.1021/acsnano.8b09683

Figure Lengend Snippet: Characterization of Rev-CD encapsulated PP7-PP7-OVA particles. (A) Electrophoretic analysis: purified particles showing PP7-PP7-OVA1, PP7-PP7-OVA2, and Rev-CD bands. (B) TEM; images are indistinguishable from those of PP7-PP7-OVA1 and PP7-PP7-OVA2 without enzyme packaged. (C) Kinetic analysis, plotted on a per-enzyme basis, of the conversion of 5-FC to 5-FU catalyzed by the indicated forms of cytosine deaminase. Solid curves show the best fit using the Michaelis–Menten equation. The @ symbol designates the encapsidated enzyme.

Article Snippet: The PP7 VLPs were expressed and assembled in BL21(DE3) E. coli cells (Biogen) as previously described for Q β .

Techniques: Purification