A conserved folding nucleus sculpts the free energy landscape of bacterial and archaeal orthologs from a divergent TIM barrel family
Harms, Michael J.
Shaffer, Scott A.
Matthews, C. Robert
UMass Chan AffiliationsThe Mass Spectrometry Facility
Department of Biochemistry and Molecular Pharmacology
Document TypeJournal Article
KeywordsTIM barrel orthologs
hydrogen deuterium exchange
Amino Acids, Peptides, and Proteins
Enzymes and Coenzymes
MetadataShow full item record
AbstractThe amino acid sequences of proteins have evolved over billions of years, preserving their structures and functions while responding to evolutionary forces. Are there conserved sequence and structural elements that preserve the protein folding mechanisms? The functionally diverse and ancient (betaalpha)1-8 TIM barrel motif may answer this question. We mapped the complex six-state folding free energy surface of a approximately 3.6 billion y old, bacterial indole-3-glycerol phosphate synthase (IGPS) TIM barrel enzyme by equilibrium and kinetic hydrogen-deuterium exchange mass spectrometry (HDX-MS). HDX-MS on the intact protein reported exchange in the native basin and the presence of two thermodynamically distinct on- and off-pathway intermediates in slow but dynamic equilibrium with each other. Proteolysis revealed protection in a small (alpha1beta2) and a large cluster (beta5alpha5beta6alpha6beta7) and that these clusters form cores of stability in Ia and Ibp The strongest protection in both states resides in beta4alpha4 with the highest density of branched aliphatic side chain contacts in the folded structure. Similar correlations were observed previously for an evolutionarily distinct archaeal IGPS, emphasizing a key role for hydrophobicity in stabilizing common high-energy folding intermediates. A bioinformatics analysis of IGPS sequences from the three superkingdoms revealed an exceedingly high hydrophobicity and surprising alpha-helix propensity for beta4, preceded by a highly conserved betaalpha-hairpin clamp that links beta3 and beta4. The conservation of the folding mechanisms for archaeal and bacterial IGPS proteins reflects the conservation of key elements of sequence and structure that first appeared in the last universal common ancestor of these ancient proteins.
Jain R, Muneeruddin K, Anderson J, Harms MJ, Shaffer SA, Matthews CR. A conserved folding nucleus sculpts the free energy landscape of bacterial and archaeal orthologs from a divergent TIM barrel family. Proc Natl Acad Sci U S A. 2021 Apr 27;118(17):e2019571118. doi: 10.1073/pnas.2019571118. PMID: 33875592; PMCID: PMC8092565. Link to article on publisher's site
Permanent Link to this Itemhttp://hdl.handle.net/20.500.14038/41901
RightsCopyright © 2021 the Author(s). Published by PNAS. This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY).
Except where otherwise noted, this item's license is described as Copyright © 2021 the Author(s). Published by PNAS. This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY).