Characterization of Recombinant Protein Mutants by Top-down Sequencing Using Quadrupole Time-of-Flight Mass Spectrometry

2009 ◽  
Vol 15 (5) ◽  
pp. 641-649 ◽  
Author(s):  
Deepalakshmi Dakshinamoorthy Putchen
Keyword(s):  
Mass Spectrometry ◽  
Recombinant Proteins ◽  
Acyl Carrier Protein ◽  
Time Of Flight ◽  
Biologically Active ◽  
Fragmentation Pattern ◽  
Post Translational Modification ◽  
Top Down ◽  
E Coli ◽  
Flight Mass Spectrometry

Top-down sequencing using quadrupole time-of-flight mass spectrometry is used as a direct way of locating the mutated sites of recombinant proteins and post-translational modification in a protein. Several mutants of barstar, expressed in E. coli, were confirmed by analyzing the fragmentation pattern of mutants. A contaminant protein, that appeared while purifying mutants of barstar, was identified as an acyl carrier protein from E. coli with a post-translational modification on serine residue, indicating that the protein was biologically active. A mutant of ribosomal protein S6 has been characterized with neutral loss of ammonia at the N-terminal region of the protein. The power of the top-down approach in characterizing the mutants of recombinant proteins has been demonstrated.

Top-down N-terminal sequencing of Immunoglobulin subunits with electrospray ionization time of flight mass spectrometry

2009 ◽  
Vol 384 (1) ◽  
pp. 42-48 ◽  
Author(s):  
Da Ren ◽  
Gary D. Pipes ◽  
David Hambly ◽  
Pavel V. Bondarenko ◽  
Michael J. Treuheit ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Electrospray Ionization ◽  
Time Of Flight ◽  
Top Down ◽  
Ionization Time ◽  
Flight Mass Spectrometry

Infrared Multiphoton Dissociation in Quadrupole Time-of-Flight Mass Spectrometry:  Top-Down Characterization of Proteins

2006 ◽  
Vol 78 (13) ◽  
pp. 4572-4577 ◽  
Author(s):  
Serguei A. Raspopov ◽  
Ayman El-Faramawy ◽  
Bruce A. Thomson ◽  
K. W. Michael Siu
Keyword(s):  
Mass Spectrometry ◽  
Time Of Flight ◽  
Infrared Multiphoton Dissociation ◽  
Top Down ◽  
Flight Mass Spectrometry ◽  
Multiphoton Dissociation

scholarly journals Identification of Polyamide 6 and Polyamide 66 Oligomers Using Ion Mobility Time-of-flight Mass Spectrometry and Risk Assessment Study of Their Migration from Kitchenware to Foodstuff

2020 ◽  
Vol 8 ◽  
Author(s):  
Xuechao Song ◽  
Elena Canellas ◽  
Cristina Nerín
Keyword(s):  
Mass Spectrometry ◽  
Ion Mobility ◽  
Extraction Method ◽  
Time Of Flight ◽  
Polyamide 6 ◽  
Fragmentation Pattern ◽  
Polyamide 66 ◽  
Flight Mass Spectrometry ◽  
Assessment Study ◽  
Whole Milk

Ion mobility time-of-flight mass spectrometry has been used for the identification the PA 6 and PA 66 oligomers through their accurate m/z, CCS values and fragmentation pattern. An extraction method for the oligomers from sunflower oil, cooked beans, soup and whole milk has been developed with recoveries ranging from 87 to 102 %.

scholarly journals Top-down analysis of 30–80 kDa proteins by electron transfer dissociation time-of-flight mass spectrometry

2013 ◽  
Vol 405 (26) ◽  
pp. 8505-8514 ◽  
Author(s):  
Luca Fornelli ◽  
Julien Parra ◽  
Ralf Hartmer ◽  
Carsten Stoermer ◽  
Markus Lubeck ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Electron Transfer ◽  
Electron Transfer Dissociation ◽  
Time Of Flight ◽  
Top Down ◽  
Flight Mass Spectrometry

scholarly journals Alkaloids Profiling of Fumaria capreolata by Analytical Platforms Based on the Hyphenation of Gas Chromatography and Liquid Chromatography with Quadrupole-Time-of-Flight Mass Spectrometry

2017 ◽  
Vol 2017 ◽  
pp. 1-16 ◽  
Author(s):  
María del Mar Contreras ◽  
Noureddine Bribi ◽  
Ana María Gómez-Caravaca ◽  
Julio Gálvez ◽  
Antonio Segura-Carretero
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Liquid Chromatography ◽  
Analytical Method ◽  
Time Of Flight ◽  
Reversed Phase ◽  
Fragmentation Pattern ◽  
Flight Mass Spectrometry ◽  
Fumaria Capreolata ◽  
Analytical Platforms

Two analytical platforms, gas chromatography (GC) coupled to quadrupole-time-of-flight (QTOF) mass spectrometry (MS) and reversed-phase ultrahigh performance liquid chromatography (UHPLC) coupled to diode array (DAD) and QTOF detection, were applied in order to study the alkaloid profile of Fumaria capreolata. The use of these mass analyzers enabled tentatively identifying the alkaloids by matching their accurate mass signals and suggested molecular formulae with those previously reported in libraries and databases. Moreover, the proposed structures were corroborated by studying their fragmentation pattern obtained by both platforms. In this way, 8 and 26 isoquinoline alkaloids were characterized using GC-QTOF-MS and RP-UHPLC-DAD-QTOF-MS, respectively, and they belonged to the following subclasses: protoberberine, protopine, aporphine, benzophenanthridine, spirobenzylisoquinoline, morphinandienone, and benzylisoquinoline. Moreover, the latter analytical method was selected to determine at 280 nm the concentration of protopine (9.6 ± 0.7 mg/g), a potential active compound of the extract. In conclusion, although GC-MS has been commonly used for the analysis of this type of phytochemicals, RP-UHPLC-DAD-QTOF-MS provided essential complementary information. This analytical method can be applied for the quality control of phytopharmaceuticals containing Fumaria extracts currently found in the market.

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