TY - JOUR
T1 - Quantitative characterization of all single amino acid variants of a viral capsid-based drug delivery vehicle
AU - Hartman, Emily C.
AU - Jakobson, Christopher M.
AU - Favor, Andrew H.
AU - Lobba, Marco J.
AU - Álvarez-Benedicto, Ester
AU - Francis, Matthew B.
AU - Tullman-Ercek, Danielle
N1 - Funding Information:
This work was funded by the Army Research Office (W911NF-15-1-0144 and W911NF-16-1-0169) and the BASF CARA program at UC Berkeley. E.C.H. was supported under by the DoD, Air Force Office of Scientific Research, National Defense Science and Engineering Graduate (NDSEG) Fellowship, 32 CFR 168a. E.A.B. was supported under the NIH-MARC Program undergraduate fellowship (5T34GM007821-38) and the Amgen Scholars Foundation. We would like to thank Dr. Ke Bi in the Computational Genomics Research Facility at UC Berkeley and Han Teng Wong for helpful discussions. The sequencing was carried by the DNA Technologies and Expression Analysis Cores at the UC Davis Genome Center, supported by NIH Shared Instrumentation Grant S10 OD010786.
Publisher Copyright:
© 2018 The Author(s).
PY - 2018/12/1
Y1 - 2018/12/1
N2 - Self-assembling proteins are critical to biological systems and industrial technologies, but predicting how mutations affect self-assembly remains a significant challenge. Here, we report a technique, termed SyMAPS (Systematic Mutation and Assembled Particle Selection), that can be used to characterize the assembly competency of all single amino acid variants of a self-assembling viral structural protein. SyMAPS studies on the MS2 bacteriophage coat protein revealed a high-resolution fitness landscape that challenges some conventional assumptions of protein engineering. An additional round of selection identified a previously unknown variant (CP[T71H]) that is stable at neutral pH but less tolerant to acidic conditions than the wild-type coat protein. The capsids formed by this variant could be more amenable to disassembly in late endosomes or early lysosomes - a feature that is advantageous for delivery applications. In addition to providing a mutability blueprint for virus-like particles, SyMAPS can be readily applied to other self-assembling proteins.
AB - Self-assembling proteins are critical to biological systems and industrial technologies, but predicting how mutations affect self-assembly remains a significant challenge. Here, we report a technique, termed SyMAPS (Systematic Mutation and Assembled Particle Selection), that can be used to characterize the assembly competency of all single amino acid variants of a self-assembling viral structural protein. SyMAPS studies on the MS2 bacteriophage coat protein revealed a high-resolution fitness landscape that challenges some conventional assumptions of protein engineering. An additional round of selection identified a previously unknown variant (CP[T71H]) that is stable at neutral pH but less tolerant to acidic conditions than the wild-type coat protein. The capsids formed by this variant could be more amenable to disassembly in late endosomes or early lysosomes - a feature that is advantageous for delivery applications. In addition to providing a mutability blueprint for virus-like particles, SyMAPS can be readily applied to other self-assembling proteins.
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U2 - 10.1038/s41467-018-03783-y
DO - 10.1038/s41467-018-03783-y
M3 - Article
C2 - 29643335
AN - SCOPUS:85045401553
SN - 2041-1723
VL - 9
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 1385
ER -