Development of a peptide probe for the occurrence of hydrogen (1H/2H) scrambling upon gas-phase fragmentation
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Development of a peptide probe for the occurrence of hydrogen (1H/2H) scrambling upon gas-phase fragmentation. / Rand, Kasper Dyrberg; Jørgensen, Thomas.
In: Analytical Chemistry, Vol. 79, No. 22, 2007, p. 8686-93.Research output: Contribution to journal › Journal article › Research › peer-review
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T1 - Development of a peptide probe for the occurrence of hydrogen (1H/2H) scrambling upon gas-phase fragmentation
AU - Rand, Kasper Dyrberg
AU - Jørgensen, Thomas
PY - 2007
Y1 - 2007
N2 - Hydrogen (1H/2H) exchange combined with mass spectrometry (HX-MS) has become a valuable method for the analysis of protein structural dynamics. Currently, localization of the incorporated deuterons is made by enzymatic cleavage of the labeled proteins, and single-residue resolution is typically only achieved for a few residues. Determination of site-specific deuterium levels by gas-phase fragmentation would greatly increase the applicability of the HX-MS method. It is, however, mandatory for this gas-phase approach that hydrogen (1H/2H) scrambling in the gaseous peptide is negligible. Thus, it is important to have a simple reference system where the onset of scrambling processes is readily detected. Here we describe a simple well-characterized set of peptides with a unique regioselective labeling that ensures an inherent high sensitivity for the detection of scrambling. This selective labeling is achieved by utilizing differences in the intrinsic exchange rates between various amino acid residues. We demonstrate that our peptides can be infused directly into an electrospray ion source by means of a cooled glass syringe, while maintaining their selective labeling in solution. We further show that the selective labeling is completely erased upon low-energy collisional activation in a tandem mass spectrometry experiment as a result of extensive hydrogen (1H/2H) scrambling.
AB - Hydrogen (1H/2H) exchange combined with mass spectrometry (HX-MS) has become a valuable method for the analysis of protein structural dynamics. Currently, localization of the incorporated deuterons is made by enzymatic cleavage of the labeled proteins, and single-residue resolution is typically only achieved for a few residues. Determination of site-specific deuterium levels by gas-phase fragmentation would greatly increase the applicability of the HX-MS method. It is, however, mandatory for this gas-phase approach that hydrogen (1H/2H) scrambling in the gaseous peptide is negligible. Thus, it is important to have a simple reference system where the onset of scrambling processes is readily detected. Here we describe a simple well-characterized set of peptides with a unique regioselective labeling that ensures an inherent high sensitivity for the detection of scrambling. This selective labeling is achieved by utilizing differences in the intrinsic exchange rates between various amino acid residues. We demonstrate that our peptides can be infused directly into an electrospray ion source by means of a cooled glass syringe, while maintaining their selective labeling in solution. We further show that the selective labeling is completely erased upon low-energy collisional activation in a tandem mass spectrometry experiment as a result of extensive hydrogen (1H/2H) scrambling.
U2 - 10.1021/ac0710782
DO - 10.1021/ac0710782
M3 - Journal article
C2 - 17935303
VL - 79
SP - 8686
EP - 8693
JO - Industrial And Engineering Chemistry Analytical Edition
JF - Industrial And Engineering Chemistry Analytical Edition
SN - 0003-2700
IS - 22
ER -
ID: 40129953