Confirmation of intersubunit connectivity and topology of designed protein complexes by native MS.
Aniruddha Sahasrabuddhe,Yang Hsia,Florian Busch,William Sheffler,Neil P. King,David Baker,Vicki H. Wysocki +6 more
TLDR
This work designs protein–protein interfaces between dimers and trimers to generate dodecameric protein assemblies with dihedral point group symmetry and shows that information about stoichiometry, intersubunit connectivity, and complex topology is rapidly gained by this SID-IM-MS methodology.Abstract:
Computational protein design provides the tools to expand the diversity of protein complexes beyond those found in nature. Understanding the rules that drive proteins to interact with each other enables the design of protein–protein interactions to generate specific protein assemblies. In this work, we designed protein–protein interfaces between dimers and trimers to generate dodecameric protein assemblies with dihedral point group symmetry. We subsequently analyzed the designed protein complexes by native MS. We show that the use of ion mobility MS in combination with surface-induced dissociation (SID) allows for the rapid determination of the stoichiometry and topology of designed complexes. The information collected along with the speed of data acquisition and processing make SID ion mobility MS well-suited to determine key structural features of designed protein complexes, thereby circumventing the requirement for more time- and sample-consuming structural biology approaches.read more
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Journal ArticleDOI
Programmable design of orthogonal protein heterodimers
Zibo Chen,Scott E. Boyken,Mengxuan Jia,Florian Busch,David Flores-Solis,Matthew J. Bick,Peilong Lu,Zachary L. VanAernum,Aniruddha Sahasrabuddhe,Robert A. Langan,Sherry Bermeo,T. J. Brunette,Vikram Khipple Mulligan,Lauren Carter,Frank DiMaio,Nikolaos G. Sgourakis,Vicki H. Wysocki,David Baker +17 more
TL;DR: The ability to design orthogonal protein heterodimers should enable sophisticated protein-based control logic for synthetic biology, and illustrates that nature has not fully explored the possibilities for programmable biomolecular interaction modalities.
Journal ArticleDOI
High-Resolution Native Mass Spectrometry.
TL;DR: A review of recent developments in high-resolution native mass spectrometer applications can be found in this article, which describes applications in the structural analysis of protein assemblies, proteoform profiling of biopharmaceuticals and plasma proteins.
Journal ArticleDOI
Controlling protein assembly on inorganic crystals through designed protein interfaces.
TL;DR: This work designs proteins displaying arrays of up to 54 carboxylate residues geometrically matched to the potassium ion (K+) sublattice on muscovite mica, demonstrating that protein–inorganic lattice interactions can be systematically programmed and set the stage for designing protein-inorganic hybrid materials.
Journal ArticleDOI
Surface-Induced Dissociation: An Effective Method for Characterization of Protein Quaternary Structure.
Alyssa Q. Stiving,Zachary L. VanAernum,Florian Busch,Sophie R. Harvey,Samantha Sarni,Vicki H. Wysocki +5 more
TL;DR: The major application focused on in this review is the structural characterization of native protein complexes, complexes kinetically trapped that retain native-like solution structures upon transfer to the gas-phase and throughout the relatively short timeframe of the mass spectrometry experiment.
Journal ArticleDOI
Higher-order structural characterisation of native proteins and complexes by top-down mass spectrometry.
Mowei Zhou,Carter Lantz,Kyle A. Brown,Ying Ge,Ljiljana Paša-Tolić,Joseph A. Loo,Frederik Lermyte +6 more
TL;DR: While this work is written primarily from a mass spectrometry perspective, it is targeted to all bioanalytical scientists who are interested in applying these methods to their own biochemistry and chemical biology research.
References
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