Singlet oxygen‐induced protein aggregation: Lysozyme crosslink formation and nLC‐MS/MS characterization

Singlet molecular oxygen (1O2) has been associated with a number of physiological processes. Despite the recognized importance of 1O2‐mediated protein modifications, little is known about the role of this oxidant in crosslink formation and protein aggregation. Thus, using lysozyme as a model, the pr...

Full description

Saved in:
Bibliographic Details
Published in:Journal of mass spectrometry. 2019-11, Vol.54 (11), p.894-905
Main Authors: Marques, Emerson Finco, Medeiros, Marisa H.G., Di Mascio, Paolo
Format: Article
Language:English
Subjects:
Agglomeration
Aggregation
Alkylation
Amino acids
Amino Acids - chemistry
Chromatography, High Pressure Liquid
Cross-Linking Reagents - chemistry
crosslink
Crosslinking
Gel electrophoresis
Histidine
Ionization
Ions
Liquid chromatography
Lysine
Lysozyme
Mass spectrometry
Mass spectroscopy
Muramidase - chemistry
Organic chemistry
Oxidants
Oxidation-Reduction
Oxidizing agents
Oxygen
Photochemical Processes
protein
Protein Aggregates
Protein interaction
Proteins
singlet molecular oxygen
Singlet oxygen
Singlet Oxygen - chemistry
Sodium lauryl sulfate
Spectrometry, Mass, Electrospray Ionization
Tandem Mass Spectrometry
Tryptophan
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Singlet molecular oxygen (1O2) has been associated with a number of physiological processes. Despite the recognized importance of 1O2‐mediated protein modifications, little is known about the role of this oxidant in crosslink formation and protein aggregation. Thus, using lysozyme as a model, the present study sought to investigate the involvement of 1O2 in crosslink formation. Lysozyme was photochemically oxidized in the presence of rose bengal or chemically oxidized using [18O]‐labeled 1O2 released from thermolabile endoperoxides. It was concluded that both 1O2 generating systems induce lysozyme crosslinking and aggregation. Using SDS‐PAGE and nano‐scale liquid chromatography coupled to electrospray ionization mass spectrometry, the results clearly demonstrated that 1O2 is directly involved in the formation of covalent crosslinks involving the amino acids histidine, lysine, and tryptophan.
ISSN:1076-5174
1096-9888
DOI:10.1002/jms.4448