The development of organometallic OBOC peptide libraries and sequencing of N-terminal rhenium(I) tricarbonyl-containing peptides utilizing MALDI tandem mass spectrometry

2015 ◽  
Vol 93 (2) ◽  
pp. 234-243 ◽  
Author(s):  
Dana R. Cruickshank ◽  
Leonard G. Luyt
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Systematic Investigation ◽  
Peptide Fragmentation ◽  
Peptide Libraries ◽  
Imaging Agents ◽  
Tandem Mass ◽  
Binding Properties ◽  
Fragmentation Patterns ◽  
Large Peptide

The development of peptide-based imaging agents through screening of large peptide libraries is hindered by the additional requirement of a radionuclide−chelator complex that can negatively affect the binding properties of the peptide. Herein, we report N-terminal rhenium(I)tricarbonyl OBOC (one-bead, one-compound) peptide libraries for use in the direct screening of potential imaging agents. The rhenium(I) tricarbonyl is incorporated directly in the library as an imaging entity surrogate to account for the presence of a technetium-99m radionuclide chelate. The identification of unknown organometallic peptides on single beads is successfully accomplished through MALDI tandem mass spectrometry, preceded by a systematic investigation of the effects of a variety of N-terminal rhenium(I) tricarbonyl chelates on peptide fragmentation patterns.

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.

Data Mining in Protein Identification by Tandem Mass Spectrometry

2011 ◽  
pp. 472-478
Author(s):  
Haipeng Wang
Keyword(s):  
Mass Spectrometry ◽  
Data Mining ◽  
Tandem Mass Spectrometry ◽  
Large Scale ◽  
Protein Identification ◽  
Peptide Fragmentation ◽  
Tandem Mass ◽  
Fragmentation Pattern ◽  
Post Translational Modification ◽  
Challenges And Opportunities

Protein identification (sequencing) by tandem mass spectrometry is a fundamental technique for proteomics which studies structures and functions of proteins in large scale and acts as a complement to genomics. Analysis and interpretation of vast amounts of spectral data generated in proteomics experiments present unprecedented challenges and opportunities for data mining in areas such as data preprocessing, peptide-spectrum matching, results validation, peptide fragmentation pattern discovery and modeling, and post-translational modification (PTM) analysis. This article introduces the basic concepts and terms of protein identification and briefly reviews the state-of-the-art relevant data mining applications. It also outlines challenges and future potential hot spots in this field.

Structural Characterization of Nitrated 2′-Hydroxychalcones by Electrospray Ionization Tandem Mass Spectrometry

2009 ◽  
Vol 15 (5) ◽  
pp. 605-616 ◽  
Author(s):  
Ana I.R.N.A. Barros ◽  
Fernando M. Nunes ◽  
Cristina Barros ◽  
Artur M.S. Silva ◽  
M. Rosário M. Domingues
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Electrospray Ionization ◽  
Structural Information ◽  
Tandem Mass ◽  
Substitution Pattern ◽  
Esi Ms ◽  
Positive Mode ◽  
Fragmentation Patterns ◽  
Product Ion Spectra

Isomeric 2′-hydroxychalcones bearing nitro and methoxy groups in different positions of their skeleton were analyzed by tandem mass spectrometry (MS/MS) with electrospray ionization (ESI), in positive mode. Collision-induced dissociation of the protonated molecules, [M + H]+, formed under electrospray conditions were studied and it was found that the product ion spectra of these chalcones presented different fragmentation patterns depending on the position of the substituents on the molecule. The product ion spectra (ESI-MS/MS) of the B ring ortho-nitro substituted 2′-hydroxychalcone and of the 4′-methoxychalcones showed loss of OH•, 2OH• and combined losses of OH• and H2O. These fragment ions were absent in the spectra of the respective meta- and para isomers. The observed differences in the product ion spectra of these nitrochalcones allowed identification of the o-nitro derivatives. Distinction between the meta- and para-derivatives was not achieved. Chalcones bearing 6′-methoxy substituents showed distinct fragmentation from the one observed for their isomers, 4′-methoxychalcones, since they present only one fragment ion, a typical (0,αA – H)+ and, therefore, do not allow detailed structural information to be obtained, nor to differentiate between the o-, m- or p-nitro isomers. Overall, it was found that small changes in the substitution pattern of chalcones change their fragmentation considerably in the ESI-MS/MS, and that these features permit the differentiation of specific isomers of these 2′-hydroxynitrochalcones.

Mining a Tandem Mass Spectrometry Database To Determine the Trends and Global Factors Influencing Peptide Fragmentation

2003 ◽  
Vol 75 (22) ◽  
pp. 6251-6264 ◽  
Author(s):  
Eugene A. Kapp ◽  
Frédéric Schütz ◽  
Gavin E. Reid ◽  
James S. Eddes ◽  
Robert L. Moritz ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Peptide Fragmentation ◽  
Tandem Mass ◽  
Factors Influencing

Characterization of the prime and non-prime active site specificities of proteases by proteome-derived peptide libraries and tandem mass spectrometry

2011 ◽  
Vol 6 (1) ◽  
pp. 111-120 ◽  
Author(s):  
Oliver Schilling ◽  
Pitter F Huesgen ◽  
Olivier Barré ◽  
Ulrich auf dem Keller ◽  
Christopher M Overall
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Active Site ◽  
Peptide Libraries ◽  
Tandem Mass
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