An Electrospray Ionization−Flow Tube Study of H/D Exchange in Protonated Bradykinin

Levy-Seri, Elchanan; Koster, Grielof; Kogan, Alexandra; Gutman, Karnit; Reuben, Bryan G.; Lifshitz, Chava

doi:10.1021/jp0037145

Cited by 23 publications

(31 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…They observed a shift of three mass units for the y 6 and y 7 ions, and two mass units for the b 6 ion. Also, they report the neutral loss of water from the precursor ion does not contain any deuterium [24]. However, they also report any conclusions based on these data are tentative because the ionizing proton at the amino terminus is expected to be mobile (assuming a salt-bridge structure), thereby causing deuterium scrambling upon ion activation [24].…”

Section: Introductionmentioning

confidence: 99%

“…The ion mobility and H/D exchange of the nonapeptide bradykinin and related analogues have been extensively studied [10,12,[15][16][17][18][19][20][21][22][23][24][25][26][27][28]. Ion mobility measurements have shown that the singly-and doubly-protonated ions of bradykinin have a similar cross-section, while the triplyprotonated ion has a cross-section that is approximately 15% larger, providing evidence for a much less compact structure [15][16][17].…”

Section: Introductionmentioning

confidence: 99%

“…Ion mobility measurements have shown that the singly-and doubly-protonated ions of bradykinin have a similar cross-section, while the triplyprotonated ion has a cross-section that is approximately 15% larger, providing evidence for a much less compact structure [15][16][17]. H/D exchange studies with D 2 O, CD 3 OD, ND 3 , and DI have detected two non-interconverting ion populations for doubly-and triply-protonated bradykinin [20,22,[24][25][26][27]. These ion populations were not detected by traditional ion mobility experiments [15][16][17], suggesting the conformers have a similar collisional cross-section.…”

Section: Introductionmentioning

confidence: 99%

“…The bradykinin ions may be folding or unfolding on the longer timescale of the FT-ICR during accumulation in the hexapole or during the H/D exchange experiment. Calculation of site-specific rate constants by Lifshitz and co-workers revealed the first three exchanges of doubly-protonated bradykinin with CD 3 OD and ND 3 have equivalent rate constants [24]. They attribute these first three exchanges to the hydrogens at the protonated amino terminus.…”

Section: Introductionmentioning

confidence: 99%

“…Also, the slower-reacting population could be converted back to the mixture of conformations through gentle activation [26]. Lifshitz and co-workers attempted to fragment doubly-protonated bradykinin labeled via H/D exchange with ND 3 [24]. They observed a shift of three mass units for the y 6 and y 7 ions, and two mass units for the b 6 ion.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Fragmentation of doubly-protonated peptide ion populations labeled by H/D exchange with CD3OD

Herrmann

Kuppannan

Wysocki

2006

International Journal of Mass Spectrometry

View full text Add to dashboard Cite

Doubly-protonated bradykinin (RPPGFSPFR) and an angiotensin III analogue (RVYIFPF) were subjected to hydrogen/deuterium (H/D) exchange with CD 3 OD in a Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer. A bimodal distribution of deuterium incorporation was present for bradykinin after H/D exchange for 90 s at a CD 3 OD pressure of 4 × 10 −7 Torr, indicating the existence of at least two distinct populations. Bradykinin ion populations corresponding to 0-2 and 5-11 deuteriums (i.e., D 0 , D 1 , D 2 , D 5 , D 6 , D 7 , D 8 , D 9 , D 10 , and D 11 ) were each monoisotopically selected and fragmented via sustained off-resonance irradiation (SORI) collision-induced dissociation (CID). The D 0 -D 2 ion populations, which correspond to the slower exchanging population, consistently require lower SORI amplitude to achieve a similar precursor ion survival yield as the faster-reacting (D 5 -D 11 ) populations. These results demonstrate that conformation/ protonation motif has an effect on fragmentation efficiency for bradykinin. Also, the partitioning of the deuterium atoms into fragment ions suggests that the C-terminal arginine residue exchanges more rapidly than the N-terminal arginine. Total deuterium incorporation in the b 1 /y 8 and b 2 /y 7 ion pairs matches very closely the theoretical values for all ion populations studied, indicating that the ions of a complementary pair are likely formed during the same fragmentation event, or that no scrambling occurs upon SORI. Deuterium incorporation into the y 1 /a 8 pseudo-ion pair does not closely match the expected theoretical values. The other peptide, doubly-protonated RVYIFPF, has a trimodal distribution of deuterium incorporation upon H/D exchange with CD 3 OD at a pressure of 1 × 10 −7 Torr for 600 s, indicating at least three distinct ion populations. After 90 s of H/D exchange where at least two distinct populations are detected, the D 0 -D 7 ion populations were monoisotopically selected and fragmented via SORI-CID over a range of SORI amplitudes. The precursor ion survival yield as a function of SORI amplitude falls into two distinct behaviors corresponding to slower-and faster-reacting ion populations. The slower-reacting population requires larger SORI amplitudes to achieve the same precursor ion survival yield as the faster exchanging population. Total deuterium incorporation into the y 2 /b 5 ion pairs matches closely the theoretical values over all ion populations and SORI amplitudes studied. This result indicates the y 2 and b 5 ions are likely formed by the same mechanism over the SORI amplitudes studied.

show abstract

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Fragmentation of doubly-protonated peptide ion populations labeled by H/D exchange with CD3OD

Herrmann

Kuppannan

Wysocki

2006

International Journal of Mass Spectrometry

View full text Add to dashboard Cite

show abstract

Charge state effect on the zwitterion influence on stability of non‐covalent interaction of single‐stranded DNA with peptides

2007

View full text Add to dashboard Cite

Negative ion ESI mass spectrometry was used to study the gas-phase stability and dissociation pathways of peptide-DNA complexes. We show that bradykinin and three modified peptides containing the basic residue arginine or lysine form stable interactions with single-stranded oligonucleotides. ESI-MS/MS of complexes of T(8) with PPGFSPFRR resulted in a major dissociation pathway through cleavage of the peptide covalent bond. The stability of the complex is due to electrostatic interaction between the negatively charged phosphate group and the basic side chain of the arginine and lysine residues as demonstrated by Vertes et al. and Woods et al. In fact, the present work establishes the role played by zwitterions on complex stabilisation. The presence of protons in nucleobase and/or amino acid contributes in reinforcing the strength of the salt bridge (SB) interaction. The zwitterionic form of the most basic of amino acid residues, arginine, is assumed to form a strong SB interaction to the negatively charged phosphate groups of DNA. This non-covalent complex is stable enough to withstand disruption of the non-covalent interaction and to first break the covalent bond. Moreover, the dependence of fragmentation patterns upon the complex charge state is explained by the fact that the net number of negative charges modulates the number of zwitterionic sites, which stabilise the complexes. Finally, the weak influence of the nucleobase is assumed by the existence of competition for proton addition between the nucleobase and the R/K side chain leading to a decrease in the stabilisation of the SB interaction.

show abstract

An electrospray‐ionization—flow‐tube study of H/D exchange in protonated leucine‐enkephalin

Ustyuzhanin

Kogan

Reuben

et al. 2001

Int J of Chemical Kinetics

Self Cite

View full text Add to dashboard Cite

An electrospray ionization fast flow technique has been employed to study reactions with CH 3 OD and ND 3 , respectively, of the protonated pentapeptide leucine enkephalin, and of protonated tri-and tetrapeptides that make up its backbone structure. Deconvolution of the experimental mass spectral data followed by simulation of the kinetic data by solution of differential equations leads to sets of apparent and site-specific rate constants. On a time scale of several milliseconds, leucine enkephalin undergoes with ND 3 four fast H/D exchanges and one slow exchange. Three equivalent exchanges are observed that are ascribed to the protonated amine group at the N-terminus. Contrary to leucine enkephalin, some of the smaller peptides that do not have tyrosine at the N-terminus are reactive with CH 3 OD, but the reactions are more than two orders of magnitude slower than the ND 3 reactions. These peptides do not sequester the ionizing proton at the N-terminus. The present study demonstrates the capabilities of a fast flow technique in studying gas phase reactions of small biomolecules. It forms part of an effort to understand gas phase conformational changes in proteins through hydrogen-exchange electrospray ionization mass spectrometry.

show abstract

An Electrospray Ionization−Flow Tube Study of H/D Exchange in Protonated Bradykinin

Cited by 23 publications

References 53 publications

Fragmentation of doubly-protonated peptide ion populations labeled by H/D exchange with CD3OD

Fragmentation of doubly-protonated peptide ion populations labeled by H/D exchange with CD3OD

Charge state effect on the zwitterion influence on stability of non‐covalent interaction of single‐stranded DNA with peptides

An electrospray‐ionization—flow‐tube study of H/D exchange in protonated leucine‐enkephalin

Contact Info

Product

Resources

About