Ion Mobility-Mass Spectrometry As a Tool for the Structural Characterization of Peptides

Ion mobility-mass spectrometry as a tool for the structural characterization of peptides bearing intra-molecular disulfide bond(s)

Philippe Massonnet(1), Jean R. N. Haler (1), Gregory Upert(2) Michel Degueldre(1) Denis Morsa(1), Nicolas Smargiasso(1), Gilles Mourier (2), Nicolas Gilles(2), Loïc Quinton(1), Edwin De Pauw*(1).

1. Laboratory of Mass Spectrometry-GIGA-R, University of Liège, Quartier Agora, Allée du six Aout 11, B-4000 Liege, Belgium
2. Commissariat à l’Energie Atomique, DRF/iBiTec – S/SIMOPRO, CE Saclay, 91191 Gif sur Yvette, France.

AUTHOR EMAIL ADDRESS:

SUPPLEMENTARY INFORMATION

SI 1) Table of the peptides used in this study.

Acm = Acetamidomethyl :

* designates a terminal amidation

O = hydroxyproline

Z = Pyroglutamic acid

Gla = Gamma carboxylic glutamic acid

SI 2) Collision Cross-Sections of peptides in different solvent conditions:

A)For peptides bearing disulfide bond(s)

B) For peptides without disulfide bond (tryptic digest)

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SI 3) Table of ion mobility calibrants used to calibrate the drift times of the travelling wave ion mobility setup.

SI 4) Calibration curve to convert drift times into Collision Cross-Sections.

-Blue linescorrespond to the 95% confidence bands

-Red lines correspond to the 95% prediction bands

SI 5) Plot of Ω(CCS) as a function of the mass-to-charge(m/z)of all the analyzed peptides at different charge states

SI 6) Plots of the CCS (Ω)as a function of the mass (m)of peptides bearing no disulfide bond atdifferent charge states (z=2 to z=6). The linear fits are depicted in black with the equations and theircorresponding R2 values.

SI 7) Plots of the CCS (Ω) as a function of the mass (m) of peptides bearing 0, 2 or 3 disulfide bonds for z=5.A linear fit function has been used to fit the trend ofthe peptides without disulfides. ∆Ω value calculations are presented in green.

SI 8)Plots of the ∆CCS (∆Ω) as a function of the mass (m) for peptides bearing different numbers of disulfide bonds (8)a) and 8)b)plots) or different cysteine connectivities (8)c) and 8)d) plots).

a)Plot of the ∆CCS (∆Ω) as a function of the mass (m) for peptides bearing different numbers of disulfide bonds for z = 4.

b)Plot of the ∆CCS (∆Ω) as a function of the mass (m) for peptides bearing different numbers of disulfide bonds for z = 5.

c) Plot of the ∆CCS (∆Ω) as a function of the mass (m) for peptides bearing different cysteine connectivities (2 intra-molecular disulfide bonds) for z = 4.

d)Plot of the ∆CCS (∆Ω) as a function of the mass (m) for peptides bearing different cysteine connectivities (2 intra-molecular disulfide bonds) for z = 5.

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