Characterization of Molecular Tweezer Binding on α-Synuclein with Native Top-Down Mass Spectrometry and Ion Mobility-Mass Spectrometry Reveals a Mechanism for Aggregation Inhibition

Lantz, Carter; Lopez, Jaybree; Goring, Andrew K.; Zenaidee, Muhammad A.; Biggs, Karl; Whitelegge, Julian P.; Ogorzalek Loo, Rachel R.; Klärner, Frank-Gerrit; Schrader, Thomas; Bitan, Gal; Loo, Joseph A.

doi:10.1021/jasms.3c00281

Cited by 2 publications

(2 citation statements)

References 62 publications

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“…Native MS, which involves analyzing proteins in aqueous solutions of high ionic strength, is an attractive alternative as non-covalent interactions may be preserved as proteins are transported to the gas phase by electrospray ionization, allowing retention of protein conformations that resemble solution structures. [31][32][33] The scope of problems that native MS can solve has expanded with the emergence of other auxiliary methods, including collision induced unfolding (CIU) and collision cross section (CCS) measurements, 33,34 variable temperature electrospray ionization (vT-ESI), [35][36][37] and alternative MS/MS methods, such as ultraviolet photodissociation (UVPD). 38 vT-ESI allows determination of melting temperatures of proteins and protein complexes 36 and measurement of thermodynamic parameters associated with ligand binding.…”

Section: Introductionmentioning

confidence: 99%

Native mass spectrometry reveals binding interactions of SARS-CoV-2 PLpro with inhibitors and ISG15

James,

Godula,

Perez

et al. 2024

Preprint

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Here we used native mass spectrometry (native MS) to probe a SARS-CoV protease, PLpro, which plays critical roles in coronavirus disease by affecting viral protein production and antagonizing host antiviral responses. Ultraviolet photodissociation (UVPD) and variable temperature electrospray ionization (vT ESI) were used to localize binding sites of PLpro inhibitors and revealed the stabilizing effects of inhibitors on protein tertiary structure. We compared PLpro from SARS-CoV-1 and SARS-CoV-2 in terms of inhibitor and ISG15 interactions to discern possible differences in protease function. A PLpro mutant lacking a single cysteine was used to localize inhibitor binding, and thermodynamic measurements revealed that inhibitor PR-619 stabilized the folded PLpro structure. These results will inform further development of PLpro as a therapeutic target against SARS-CoV-2 and other emerging coronaviruses.

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Section: Introductionmentioning

confidence: 99%

Native mass spectrometry reveals binding interactions of SARS-CoV-2 PLpro with inhibitors and ISG15

James,

Godula,

Perez

et al. 2024

Preprint

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“…Environmental factors such as post-translational modifications (PTMs), , ligand binding, − mutations, and solution conditions (pH, temperature, pressure, presence of osmolytes, and other cofactors) − can alter a protein’s complex structure, dynamics, and stability. An important factor that is often overlooked is the role of hydration in protein stability and structure.…”

Section: Introductionmentioning

confidence: 99%

Water Plays Key Roles in Stabilities of Wild Type and Mutant Transthyretin Complexes

Lantz,

Rider,

Yun

et al. 2024

J. Am. Soc. Mass Spectrom.

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Transthyretin (TTR), a 56 kDa homotetramer that is involved in the transport of thyroxine and retinol, has been linked to amyloidosis through disassembly of tetramers to form monomers, dimers, and trimers that then reassemble into higher order oligomers and/or fibrils. Hybrid TTR (hTTR) tetramers are found in heterozygous individuals that express both wild type TTR (wt-TTR) and mutant TTR (mTTR) forms of the protein, and these states display increased rates of amyloidosis. Here we monitor subunit exchange (SUE) reactions involving homomeric and mixed tetramers using high resolution native mass spectrometry (nMS). Our results show evidence that differences in TTR primary structure alter tetramer stabilities, and hTTR products can form spontaneously by SUE reactions. In addition, we find that solution temperature has strong effects on TTR tetramer stabilities and formation of SUE products. Lower temperatures promote formation of hTTR tetramers containing L55P and V30M subunits, whereas small effects on the formation of hTTR tetramers containing F87A and T119M subunits are observed. We hypothesize that the observed temperature dependent stabilities and subsequent SUE behavior are a result of perturbations to the network of "two kinds of water": hydrating and structure stabilizing water molecules (Spyrakis et al.

show abstract

Characterization of Molecular Tweezer Binding on α-Synuclein with Native Top-Down Mass Spectrometry and Ion Mobility-Mass Spectrometry Reveals a Mechanism for Aggregation Inhibition

Cited by 2 publications

References 62 publications

Native mass spectrometry reveals binding interactions of SARS-CoV-2 PLpro with inhibitors and ISG15

Native mass spectrometry reveals binding interactions of SARS-CoV-2 PLpro with inhibitors and ISG15

Water Plays Key Roles in Stabilities of Wild Type and Mutant Transthyretin Complexes

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