Charge Engineering Reveals the Roles of Ionizable Side Chains in Electrospray Ionization Mass Spectrometry
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Charge Engineering Reveals the Roles of Ionizable Side Chains in Electrospray Ionization Mass Spectrometry. / Abramsson, Mia L.; Sahin, Cagla; Hopper, Jonathan T. S.; Branca, Rui M. M.; Danielsson, Jens; Xu, Mingming; Chandler, Shane A.; Österlund, Nicklas; Ilag, Leopold L.; Leppert, Axel; Costeira-Paulo, Joana; Lang, Lisa; Teilum, Kaare; Laganowsky, Arthur; Benesch, Justin L. P.; Oliveberg, Mikael; Robinson, Carol V.; Marklund, Erik G.; Allison, Timothy M.; Winther, Jakob R.; Landreh, Michael.
In: JACS Au, Vol. 1, No. 12, 2021, p. 2385-2393.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Charge Engineering Reveals the Roles of Ionizable Side Chains in Electrospray Ionization Mass Spectrometry
AU - Abramsson, Mia L.
AU - Sahin, Cagla
AU - Hopper, Jonathan T. S.
AU - Branca, Rui M. M.
AU - Danielsson, Jens
AU - Xu, Mingming
AU - Chandler, Shane A.
AU - Österlund, Nicklas
AU - Ilag, Leopold L.
AU - Leppert, Axel
AU - Costeira-Paulo, Joana
AU - Lang, Lisa
AU - Teilum, Kaare
AU - Laganowsky, Arthur
AU - Benesch, Justin L. P.
AU - Oliveberg, Mikael
AU - Robinson, Carol V.
AU - Marklund, Erik G.
AU - Allison, Timothy M.
AU - Winther, Jakob R.
AU - Landreh, Michael
N1 - © 2021 The Authors. Published by American Chemical Society.
PY - 2021
Y1 - 2021
N2 - In solution, the charge of a protein is intricately linked to its stability, but electrospray ionization distorts this connection, potentially limiting the ability of native mass spectrometry to inform about protein structure and dynamics. How the behavior of intact proteins in the gas phase depends on the presence and distribution of ionizable surface residues has been difficult to answer because multiple chargeable sites are present in virtually all proteins. Turning to protein engineering, we show that ionizable side chains are completely dispensable for charging under native conditions, but if present, they are preferential protonation sites. The absence of ionizable side chains results in identical charge state distributions under native-like and denaturing conditions, while coexisting conformers can be distinguished using ion mobility separation. An excess of ionizable side chains, on the other hand, effectively modulates protein ion stability. In fact, moving a single ionizable group can dramatically alter the gas-phase conformation of a protein ion. We conclude that although the sum of the charges is governed solely by Coulombic terms, their locations affect the stability of the protein in the gas phase.
AB - In solution, the charge of a protein is intricately linked to its stability, but electrospray ionization distorts this connection, potentially limiting the ability of native mass spectrometry to inform about protein structure and dynamics. How the behavior of intact proteins in the gas phase depends on the presence and distribution of ionizable surface residues has been difficult to answer because multiple chargeable sites are present in virtually all proteins. Turning to protein engineering, we show that ionizable side chains are completely dispensable for charging under native conditions, but if present, they are preferential protonation sites. The absence of ionizable side chains results in identical charge state distributions under native-like and denaturing conditions, while coexisting conformers can be distinguished using ion mobility separation. An excess of ionizable side chains, on the other hand, effectively modulates protein ion stability. In fact, moving a single ionizable group can dramatically alter the gas-phase conformation of a protein ion. We conclude that although the sum of the charges is governed solely by Coulombic terms, their locations affect the stability of the protein in the gas phase.
U2 - 10.1021/jacsau.1c00458
DO - 10.1021/jacsau.1c00458
M3 - Journal article
C2 - 34977906
VL - 1
SP - 2385
EP - 2393
JO - JACS Au
JF - JACS Au
SN - 2691-3704
IS - 12
ER -
ID: 289963521