Structural studies of the sBBI/trypsin non-covalent complex using covalent modification and mass spectrometry

2014 ◽  
Vol 28 (5) ◽  
pp. 413-429 ◽  
Author(s):  
Ekaterina Darii ◽  
Guanalini Saravanamuthu ◽  
Ivo G. Gut ◽  
Jean-Claude Tabet
Keyword(s):  
Mass Spectrometry ◽  
Covalent Modification ◽  
Structural Studies ◽  
Covalent Complex

scholarly journals Characterization of a novel + 70 Da modification in rhGM-CSF expressed in E. coli using chemical assays in combination with mass spectrometry

Amino Acids ◽  
2021 ◽  
Author(s):  
Magdalena Widgren Sandberg ◽  
Jakob Bunkenborg ◽  
Stine Thyssen ◽  
Martin Villadsen ◽  
Thomas Kofoed
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Peptide Mapping ◽  
Covalent Modification ◽  
Peptide Fragmentation ◽  
Tandem Mass ◽  
E Coli ◽  
Gm Csf ◽  
Fragmentation Behavior ◽  
Macrophage Colony

AbstractGranulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine and a white blood cell growth factor that has found usage as a therapeutic protein. During analysis of different fermentation batches of GM-CSF recombinantly expressed in E. coli, a covalent modification was identified on the protein by intact mass spectrometry. The modification gave a mass shift of + 70 Da and peptide mapping analysis demonstrated that it located to the protein N-terminus and lysine side chains. The chemical composition of C4H6O was found to be the best candidate by peptide fragmentation using tandem mass spectrometry. The modification likely contains a carbonyl group, since the mass of the modification increased by 2 Da by reduction with borane pyridine complex and it reacted with 2,4-dinitrophenylhydrazine. On the basis of chemical and tandem mass spectrometry fragmentation behavior, the modification could be attributed to crotonaldehyde, a reactive compound formed during lipid peroxidation. A low recorded oxygen pressure in the reactor during protein expression could be linked to the formation of this compound. This study shows the importance of maintaining full control over all reaction parameters during recombinant protein production.

The [C5H8]+˙ radical cation: Structural studies by energy-resolved mass spectrometry

1985 ◽  
Vol 20 (12) ◽  
pp. 727-732 ◽  
Author(s):  
R. S. Mason ◽  
K. R. Jennings ◽  
S. Verma ◽  
R. G. Cooks
Keyword(s):  
Mass Spectrometry ◽  
Radical Cation ◽  
Structural Studies

Accelerator mass spectrometry for assaying irreversible covalent modification of an enzyme by acetoacetic ester

1998 ◽  
Vol 179-180 ◽  
pp. 185-193 ◽  
Author(s):  
Jacqueline S. Bennett ◽  
Darren W. Bell ◽  
Bruce A. Buchholz ◽  
Eric S.C. Kwok ◽  
John S. Vogel ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Accelerator Mass Spectrometry ◽  
Covalent Modification ◽  
Acetoacetic Ester

scholarly journals Structural evidence for the covalent modification of FabH by 4,5-dichloro-1,2-dithiol-3-one (HR45)

2017 ◽  
Vol 15 (30) ◽  
pp. 6310-6313 ◽  
Author(s):  
Alexander G. Ekström ◽  
Van Kelly ◽  
Jon Marles-Wright ◽  
Scott L. Cockroft ◽  
Dominic J. Campopiano
Keyword(s):  
Mass Spectrometry ◽  
Reaction Mechanism ◽  
Active Site ◽  
Covalent Modification ◽  
Elimination Reaction ◽  
Covalent Adduct ◽  
Michael Type Addition

Mass spectrometry and modelling shows the antimicrobial inhibitor 4,5-dichloro-1,2-dithiol-3-one (HR45) acts by forming a covalent adduct with the target β-ketoacyl-ACP synthase III (FabH). The 5-chloro substituent directs attack of the essential active site thiol (C112) via a Michael type addition elimination reaction mechanism.

Applications of mass spectrometry in structural studies of bioactive compounds

1979 ◽  
Vol 293 (1400) ◽  
pp. 3-11
Keyword(s):  
Mass Spectrometry ◽  
Cell Proliferation ◽  
Tissue Culture ◽  
Nucleic Acids ◽  
Bioactive Compounds ◽  
Rat Liver ◽  
Structural Studies

The paper describes the role played by mass spectrometry in structural studies of molluscicidal saponins from a plant; the adducts formed between 7,12-dimethylbenz[a]anthracene and nucleic acids upon tissue culture with rat liver; and the G2 factor, the first natural mitotic hormone that regulates cell proliferation.

Fast atom bombardment combined with tandem mass spectrometry for structural studies of substituted cyclic perfluoroalkanesulfonates

1987 ◽  
Vol 59 (22) ◽  
pp. 2653-2658 ◽  
Author(s):  
Sharon L. Hunt ◽  
Philip A. Lyon ◽  
Fred E. Behr ◽  
Larry D. Winter ◽  
Ronald L. Cerny ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Fast Atom Bombardment ◽  
Tandem Mass ◽  
Structural Studies

scholarly journals Diazirine-functionalized mannosides for photoaffinity labeling: trouble with FimH

2018 ◽  
Vol 14 ◽  
pp. 1890-1900 ◽  
Author(s):  
Femke Beiroth ◽  
Tomas Koudelka ◽  
Thorsten Overath ◽  
Stefan D Knight ◽  
Andreas Tholey ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Performance ◽  
Photoaffinity Labeling ◽  
Extensive Study ◽  
Covalent Modification ◽  
Bacterial Protein ◽  
High Affinity ◽  
Limited Success ◽  
Receptor Interactions ◽  
Interdisciplinary Methodology

Photoaffinity labeling is frequently employed for the investigation of ligand–receptor interactions in solution. We have employed an interdisciplinary methodology to achieve facile photolabeling of the lectin FimH, which is a bacterial protein, crucial for adhesion, colonization and infection. Following our earlier work, we have here designed and synthesized diazirine-functionalized mannosides as high-affinity FimH ligands and performed an extensive study on photo-crosslinking of the best ligand (mannoside 3) with a series of model peptides and FimH. Notably, we have employed high-performance mass spectrometry to be able to detect radiation results with the highest possible accuracy. We are concluding from this study that photolabeling of FimH with sugar diazirines has only very limited success and cannot be regarded a facile approach for covalent modification of FimH.

Sign in / Sign up

Export Citation Format

Share Document