Simple Atmospheric Hydrogen/Deuterium Exchange Method for Enumeration of Labile Hydrogens by Electrospray Ionization Mass Spectrometry

Kostyukevich, Yury; Kononikhin, Alexey; Popov, Igor; Nikolaev, E. N.

doi:10.1021/ac4006606

Cited by 83 publications

(104 citation statements)

References 9 publications

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“…All experiments were repeated at least three times, and the results were found to be reproducible. The effect of the position of the copper plate, ESI needle and the amount of the D 2 O was addressed in our previous papers …”

Section: Methodsmentioning

confidence: 99%

In ESI-source H/D exchange under atmospheric pressure for peptides and proteins of different molecular weights from 1 to 66 kDa: the role of the temperature of the desolvating capillary on H/D exchange

et al. 2015

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Transition of proteins from the solution to the gas phase during electrospray ionization remains a challenging problem despite the large amount of attention it has received during the past few decades. One of the major questions relates to the extent to which proteins in the gas phase retain their condensed phase structures. We have used in-electrospray source hydrogen/deuterium exchange to determine the number of deuterium incorporations as a function of protein mass, charge state and temperature of the desolvating capillary where the reaction occurs. All experiments were performed on a Thermo LTQ FT Ultra equipped with a 7-T superconducting magnet. Ions were generated by an IonMax Electrospray ion source operated in the positive ESI mode. Deuterium exchange was performed by introducing a droplet of D2 O beneath the ESI capillary. We systematically investigated gas phase hydrogen/deuterium (H/D) exchange under atmospheric pressure for peptides and proteins of different molecular weights from 1 to 66 kDa. We observed that almost all proteins demonstrate similar exchange rates for all charge states and that these rates increase exponentially with the temperature of the desolvating capillary. We did not observe any clear correlation of the number of H/D exchanges with the value of the cross section for a corresponding charge state. We have demonstrated the possibility of performing in-ESI source H/D exchange of large proteins under atmospheric pressure. The simplicity of the experimental setup makes it a useful experimental technique that can be applied for the investigation of gas phase conformations of proteins.

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Section: Methodsmentioning

confidence: 99%

In ESI-source H/D exchange under atmospheric pressure for peptides and proteins of different molecular weights from 1 to 66 kDa: the role of the temperature of the desolvating capillary on H/D exchange

et al. 2015

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“…More details about the used H/D exchange approach can be found elsewhere 29 . For exchange in the liquid phase, aqueous solutions of bn and b* with concentrations of 250 μM were mixed, incubated at 80 °C for 5 minutes and diluted with a deuterated buffer.…”

Section: Methodsmentioning

confidence: 99%

“…Once the complex is ionized and the 3D structure of the ion is likely to be as close as possible to obtain to the structure in the solution, then the H/D exchange reaction in the gas phase labels the outer surface of the complex and the following MS n experiments reveal the exchange-protected sites. Recently it was suggested to perform the H/D exchange reaction directly in the ESI source 28, 29 . This approach was applied to study proteins 30–32 , oligosaccharides 33 , natural complex mixtures 34–37 and small organic molecules 38 .…”

Section: Introductionmentioning

confidence: 99%

CID fragmentation, H/D exchange and supermetallization of Barnase-Barstar complex

Kostyukevich

Shulga

Kononikhin

et al. 2017

Sci Rep

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The barnase-barstar complex is one of the most stable protein-protein complexes and has a very wide range of possible applications. Here we report the use of top-down mass spectrometry for the investigation of the structure of this complex, its ionization via ESI, isolation and fragmentation. It was found that the asymmetry of the resulting charge state distributions of the protein monomer product ions increased as the charge state of the precursor ions increased. For the investigation of the 3D structure of the complex, the gas phase H/D exchange reaction was used. In addition, supermetallized ions of the complex with Zn were produced and investigated. It was observed that an increase in the number of metals bound to the complex results in a change in complex stability and the charge distribution between protein fragment. Analysis of the fragmentation pattern of the supermetallized complex [bn-b* + 5Zn]10+ indicated that this ion is present in different conformations with different charges and Zn distributions. Since Zn cannot migrate, such structures must be formed during ionization.

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“…Данная реакция позволяет делать определенные выво-ды о структуре и о конформации ионов в газовой фазе. Ранее методика проведения реакции дейтеро/водород-ного обмена в газовой фазе при атмосферном давлении прямо во время процесса ионизации [13] была успешно применена для структурного исследования широко-го класса различных соединений, таких как пепти-ды, [14,15] белки, сахара, [16,17] малые органические молеку-лы, [18][19][20][21][22] сложные природные субстанции [23,24] и олиго-нуклеотиды. [25] Результаты анализа реакции дейтеро/водородного обмена для комплексов β-циклодекстрина с Ce 3+ приведены в Таблице 1.…”

Section: результаты и обсуждениеunclassified

Investigation of Ion–Molecular Complexes of beta–Cyclodextrin with Proteins and Metals in Gas Phase

Kostyukevich¹,

Kononikhin²,

Бугрова³

et al. 2017

MHC

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Simple Atmospheric Hydrogen/Deuterium Exchange Method for Enumeration of Labile Hydrogens by Electrospray Ionization Mass Spectrometry

Cited by 83 publications

References 9 publications

In ESI-source H/D exchange under atmospheric pressure for peptides and proteins of different molecular weights from 1 to 66 kDa: the role of the temperature of the desolvating capillary on H/D exchange

In ESI-source H/D exchange under atmospheric pressure for peptides and proteins of different molecular weights from 1 to 66 kDa: the role of the temperature of the desolvating capillary on H/D exchange

CID fragmentation, H/D exchange and supermetallization of Barnase-Barstar complex

Investigation of Ion–Molecular Complexes of beta–Cyclodextrin with Proteins and Metals in Gas Phase

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