TY  - JOUR
A1  - Young, Phoebe
A1  - Hense, Genia
A1  - Immer, Carina
A1  - Wöhnert, Jens
A1  - Morgner, Nina
T1  - LILBID laser dissociation curves: a mass spectrometry-based method for the quantitative assessment of dsDNA binding affinities
T2  - Scientific reports
N2  - One current goal in native mass spectrometry is the assignment of binding affinities to noncovalent complexes. Here we introduce a novel implementation of the existing laser-induced liquid bead ion desorption (LILBID) mass spectrometry method: this new method, LILBID laser dissociation curves, assesses binding strengths quantitatively. In all LILBID applications, aqueous sample droplets are irradiated by 3 µm laser pulses. Variation of the laser energy transferred to the droplet during desorption affects the degree of complex dissociation. In LILBID laser dissociation curves, laser energy transfer is purposely varied, and a binding affinity is calculated from the resulting complex dissociation. A series of dsDNAs with different binding affinities was assessed using LILBID laser dissociation curves. The binding affinity results from the LILBID laser dissociation curves strongly correlated with the melting temperatures from UV melting curves and with dissociation constants from isothermal titration calorimetry, standard solution phase methods. LILBID laser dissociation curve data also showed good reproducibility and successfully predicted the melting temperatures and dissociation constants of three DNA sequences. LILBID laser dissociation curves are a promising native mass spectrometry binding affinity method, with reduced time and sample consumption compared to melting curves or titrations.
KW  - Biochemistry
KW  - Chemistry
KW  - Mass spectrometry
KW  - Physical chemistry
Y1  - 2020
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/79265
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-792650
SN  - 2045-2322
N1  - The ITC instrument was acquired with funds from the DFG-supported CRC 902 "Molecular Principles of RNA-based Regulation".
Open Access funding enabled and organized by Projekt DEAL.
VL  - 10
IS  - art. 20398
SP  - 1
EP  - 13
PB  - Macmillan Publishers Limited, part of Springer Nature
CY  - [London]
ER  -