Characterization and De Novo Sequencing of Atlantic Salmon Vitellogenin Protein by Electrospray Tandem and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry
Vitellogenin (VTG) is a protein produced by the liver of oviparous animals in response to circulating estrogens. In the plasma of males and immature females, VTG is undetectable. VTG has been used as a biomarker for exposure to endocrine disruptors in many species. In the present study, characteriza...
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crsagepubl:10.1255/ejms.588 2023-05-15T15:29:02+02:00 Characterization and De Novo Sequencing of Atlantic Salmon Vitellogenin Protein by Electrospray Tandem and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Banoub, Joseph Cohen, Alejandro Mansour, Atef Thibault, Pierre 2004 http://dx.doi.org/10.1255/ejms.588 http://journals.sagepub.com/doi/pdf/10.1255/ejms.588 en eng SAGE Publications http://journals.sagepub.com/page/policies/text-and-data-mining-license European Journal of Mass Spectrometry volume 10, issue 1, page 121-134 ISSN 1469-0667 1751-6838 Spectroscopy Atomic and Molecular Physics, and Optics General Medicine journal-article 2004 crsagepubl https://doi.org/10.1255/ejms.588 2022-04-14T04:40:33Z Vitellogenin (VTG) is a protein produced by the liver of oviparous animals in response to circulating estrogens. In the plasma of males and immature females, VTG is undetectable. VTG has been used as a biomarker for exposure to endocrine disruptors in many species. In the present study, characterization of intact Atlantic salmon VTG was effected using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI ToF MS). Tryptic digest peptides were analyzed by MALDI ToF MS to obtain a peptide mass fingerprint. De novo sequencing of the tryptic peptides used low-energy collisionally-induced dissociation (CID) in an electrospray ionization quadrupole-ToF orthogonal hybrid mass spectrometer (ESI Q-ToF MS/MS). The interpretation of the product-ion spectra obtained from the ESI Q-ToF MS/MS was done by Lutefisk, a computer-based software algorithm. The molecular mass of the intact protein was found to be 187335 Da. A total of 14 tryptic peptides were sequenced and compared with the complete rainbow trout VTG and the partial Atlantic salmon VTG sequences found in the Swiss-Prot database. De novo sequencing by CID MS/MS of 11 Atlantic salmon tryptic digest peptides with selected precursor ions at m/z 788.24, 700.20, 794.75, 834.31, 889.28, 819.79, 865.27, 843.81, 572.20, 573.66 and 561.68 showed high homology with the known sequence of rainbow trout VTG. The last two precursor peptide ions, found at m/z 573.66 and m/z 561.68, also specifically matched the known portion of the Atlantic salmon VTG sequence. Finally, three tryptic precursor peptide ions found at m/z 795.18, 893.28 and 791.05, provided product-ion spectra, which were exclusive to the unsequenced portion of the Atlantic salmon VTG. Article in Journal/Newspaper Atlantic salmon SAGE Publications (via Crossref) European Journal of Mass Spectrometry 10 1 121 134 |
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Open Polar |
collection |
SAGE Publications (via Crossref) |
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language |
English |
topic |
Spectroscopy Atomic and Molecular Physics, and Optics General Medicine |
spellingShingle |
Spectroscopy Atomic and Molecular Physics, and Optics General Medicine Banoub, Joseph Cohen, Alejandro Mansour, Atef Thibault, Pierre Characterization and De Novo Sequencing of Atlantic Salmon Vitellogenin Protein by Electrospray Tandem and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry |
topic_facet |
Spectroscopy Atomic and Molecular Physics, and Optics General Medicine |
description |
Vitellogenin (VTG) is a protein produced by the liver of oviparous animals in response to circulating estrogens. In the plasma of males and immature females, VTG is undetectable. VTG has been used as a biomarker for exposure to endocrine disruptors in many species. In the present study, characterization of intact Atlantic salmon VTG was effected using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI ToF MS). Tryptic digest peptides were analyzed by MALDI ToF MS to obtain a peptide mass fingerprint. De novo sequencing of the tryptic peptides used low-energy collisionally-induced dissociation (CID) in an electrospray ionization quadrupole-ToF orthogonal hybrid mass spectrometer (ESI Q-ToF MS/MS). The interpretation of the product-ion spectra obtained from the ESI Q-ToF MS/MS was done by Lutefisk, a computer-based software algorithm. The molecular mass of the intact protein was found to be 187335 Da. A total of 14 tryptic peptides were sequenced and compared with the complete rainbow trout VTG and the partial Atlantic salmon VTG sequences found in the Swiss-Prot database. De novo sequencing by CID MS/MS of 11 Atlantic salmon tryptic digest peptides with selected precursor ions at m/z 788.24, 700.20, 794.75, 834.31, 889.28, 819.79, 865.27, 843.81, 572.20, 573.66 and 561.68 showed high homology with the known sequence of rainbow trout VTG. The last two precursor peptide ions, found at m/z 573.66 and m/z 561.68, also specifically matched the known portion of the Atlantic salmon VTG sequence. Finally, three tryptic precursor peptide ions found at m/z 795.18, 893.28 and 791.05, provided product-ion spectra, which were exclusive to the unsequenced portion of the Atlantic salmon VTG. |
format |
Article in Journal/Newspaper |
author |
Banoub, Joseph Cohen, Alejandro Mansour, Atef Thibault, Pierre |
author_facet |
Banoub, Joseph Cohen, Alejandro Mansour, Atef Thibault, Pierre |
author_sort |
Banoub, Joseph |
title |
Characterization and De Novo Sequencing of Atlantic Salmon Vitellogenin Protein by Electrospray Tandem and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry |
title_short |
Characterization and De Novo Sequencing of Atlantic Salmon Vitellogenin Protein by Electrospray Tandem and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry |
title_full |
Characterization and De Novo Sequencing of Atlantic Salmon Vitellogenin Protein by Electrospray Tandem and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry |
title_fullStr |
Characterization and De Novo Sequencing of Atlantic Salmon Vitellogenin Protein by Electrospray Tandem and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry |
title_full_unstemmed |
Characterization and De Novo Sequencing of Atlantic Salmon Vitellogenin Protein by Electrospray Tandem and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry |
title_sort |
characterization and de novo sequencing of atlantic salmon vitellogenin protein by electrospray tandem and matrix-assisted laser desorption/ionization mass spectrometry |
publisher |
SAGE Publications |
publishDate |
2004 |
url |
http://dx.doi.org/10.1255/ejms.588 http://journals.sagepub.com/doi/pdf/10.1255/ejms.588 |
genre |
Atlantic salmon |
genre_facet |
Atlantic salmon |
op_source |
European Journal of Mass Spectrometry volume 10, issue 1, page 121-134 ISSN 1469-0667 1751-6838 |
op_rights |
http://journals.sagepub.com/page/policies/text-and-data-mining-license |
op_doi |
https://doi.org/10.1255/ejms.588 |
container_title |
European Journal of Mass Spectrometry |
container_volume |
10 |
container_issue |
1 |
container_start_page |
121 |
op_container_end_page |
134 |
_version_ |
1766359406509817856 |