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.
Submitter: Shu-Ching Ou
Submission Date: Dec. 20, 2018, 4:51 p.m.
|Structure ID||Release Date||Resolution||Structure Title|
|1AQ3||1997-08-06T00:00:00+0000||2.8||BACTERIOPHAGE MS2 CAPSID PROTEIN/RNA COMPLEX|
|1AQ4||1997-08-06T00:00:00+0000||3.0||STRUCTURE OF A MS2 COAT PROTEIN MUTANT IN COMPLEX WITH AN RNA OPERATOR|
|1BMS||1995-08-29T00:00:00+0000||2.7||CRYSTAL STRUCTURE OF MS2 CAPSIDS WITH MUTATIONS IN THE SUBUNIT FG LOOP|
|1MSC||1995-04-28T00:00:00+0000||2.0||CRYSTAL STRUCTURE OF MS2 COAT PROTEIN DIMER|
|1MST||1995-08-30T00:00:00+0000||2.6||CRYSTAL STRUCTURE OF MS2 CAPSIDS WITH MUTATIONS IN THE SUBUNIT FG LOOP|
|1MVA||1997-08-06T00:00:00+0000||3.0||STRUCTURE OF A PROTEIN CAPSID OF THE T45A MUTANT OF PHAGE MS2|
|1MVB||1997-08-06T00:00:00+0000||3.0||STRUCTURE OF A PROTEIN CAPSID OF THE T59S MUTANT OF PHAGE MS2|
|1U1Y||2004-07-16T00:00:00+0000||2.85||Crystal structure of a complex between WT bacteriophage MS2 coat protein and an F5 aptamer RNA stemloop with 2aminopurine substituted at the-10 position|
|1ZDH||1996-09-24T00:00:00+0000||2.7||MS2 COAT PROTEIN/RNA COMPLEX|
|1ZDI||1996-09-24T00:00:00+0000||2.7||RNA BACTERIOPHAGE MS2 COAT PROTEIN/RNA COMPLEX|