Quantitative characterization of all single amino acid variants of a viral capsid-based drug delivery vehicle.


Abstract

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.

Submission Details

ID: 2VxwaJVT

Submitter: Shu-Ching Ou

Submission Date: Dec. 20, 2018, 4:51 p.m.

Version: 1

Publication Details
Hartman EC;Jakobson CM;Favor AH;Lobba MJ;Álvarez-Benedicto E;Francis MB;Tullman-Ercek D,Nat Commun (2018) Quantitative characterization of all single amino acid variants of a viral capsid-based drug delivery vehicle. PMID:29643335
Additional Information

Number of data points 2452
Proteins Capsid protein
Unique complexes 2452
Assays/Quantities/Protocols Experimental Assay: Apparent Fitness Scores
Libraries Variants for Capsid Protein
Sequence Assay Result Units