Tandem Mass Spectrometry as Strategy for the Selective Identification and Quantification of the Amyloid Precursor Protein Tyr682 Residue Phosphorylation Status in Human Blood Mononuclear Cells
<b>Background:</b> Alzheimer’s disease (AD) is a devastating neurodegenerative disease without guidelines for early diagnosis or personalized treatment. Previous studies have highlighted a crucial role of increasing phosphorylation levels of the amyloid precursor protein (APP) Tyr682 res...
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MDPI AG
2021-08-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/11/9/1297 |
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doaj-0ebf8750d40b48c69fcee86e02d261002021-09-25T23:47:20ZengMDPI AGBiomolecules2218-273X2021-08-01111297129710.3390/biom11091297Tandem Mass Spectrometry as Strategy for the Selective Identification and Quantification of the Amyloid Precursor Protein Tyr682 Residue Phosphorylation Status in Human Blood Mononuclear CellsPierluigi Reveglia0Rosarita Nasso1Antonella Angiolillo2Lucia Lecce3Carmela Paolillo4Samantha De Tullio5Monica Gelzo6Alfonso Di Costanzo7Carmela Matrone8Gaetano Corso9Department of Clinical and Experimental Medicine, University of Foggia, 71122 Foggia, ItalyDivision of Pharmacology, Department of Neuroscience, School of Medicine, University of Naples Federico II, 80131 Naples, ItalyCentre for Research and Training in Medicine for Aging, Department of Medicine and Health Sciences “Vincenzo Tiberio”, University of Molise, 86100 Campobasso, ItalyDepartment of Clinical and Experimental Medicine, University of Foggia, 71122 Foggia, ItalyDepartment of Clinical and Experimental Medicine, University of Foggia, 71122 Foggia, ItalyDepartment of Clinical and Experimental Medicine, University of Foggia, 71122 Foggia, ItalyCEINGE-Biotecnologie Avanzate, scarl, 80145 Naples, ItalyCentre for Research and Training in Medicine for Aging, Department of Medicine and Health Sciences “Vincenzo Tiberio”, University of Molise, 86100 Campobasso, ItalyDivision of Pharmacology, Department of Neuroscience, School of Medicine, University of Naples Federico II, 80131 Naples, ItalyDepartment of Clinical and Experimental Medicine, University of Foggia, 71122 Foggia, Italy<b>Background:</b> Alzheimer’s disease (AD) is a devastating neurodegenerative disease without guidelines for early diagnosis or personalized treatment. Previous studies have highlighted a crucial role of increasing phosphorylation levels of the amyloid precursor protein (APP) Tyr682 residue in predicting neuronal deficits in AD patients. However, the lack of a method for the identification and quantification of Tyr682 phosphorylation levels prevents its potential clinical applications. <b>Methods:</b> Here we report a method to identify and quantify APP Tyr682 phosphorylation levels in blood mononuclear cells of AD patients by tandem mass spectrometry (tMS). <b>Results:</b> This method showed excellent sensitivity with detection and quantification limits set respectively at 0.035 and 0.082 ng injected for the phosphorylated peptide and at 0.02 and 0.215 ng injected for the non-phosphorylated peptide. The average levels of both peptides were quantified in transfected HELA cells (2.48 and 3.53 ng/μg of protein, respectively). Preliminary data on 3 AD patients showed quantifiable levels of phosphorylated peptide (0.10–0.15 ng/μg of protein) and below the LOQ level of non-phosphorylated peptide (0.13 ng/μg of protein). <b>Conclusion:</b> This method could allow the identification of patients with increased APP Tyr682 phosphorylation and allow early characterization of molecular changes prior to the appearance of clinical signs.https://www.mdpi.com/2218-273X/11/9/1297Alzheimer’s diseaseAPP Tyr682 phosphorylationtargeted peptide analysis |
| collection |
DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Pierluigi Reveglia Rosarita Nasso Antonella Angiolillo Lucia Lecce Carmela Paolillo Samantha De Tullio Monica Gelzo Alfonso Di Costanzo Carmela Matrone Gaetano Corso |
| spellingShingle |
Pierluigi Reveglia Rosarita Nasso Antonella Angiolillo Lucia Lecce Carmela Paolillo Samantha De Tullio Monica Gelzo Alfonso Di Costanzo Carmela Matrone Gaetano Corso Tandem Mass Spectrometry as Strategy for the Selective Identification and Quantification of the Amyloid Precursor Protein Tyr682 Residue Phosphorylation Status in Human Blood Mononuclear Cells Biomolecules Alzheimer’s disease APP Tyr682 phosphorylation targeted peptide analysis |
| author_facet |
Pierluigi Reveglia Rosarita Nasso Antonella Angiolillo Lucia Lecce Carmela Paolillo Samantha De Tullio Monica Gelzo Alfonso Di Costanzo Carmela Matrone Gaetano Corso |
| author_sort |
Pierluigi Reveglia |
| title |
Tandem Mass Spectrometry as Strategy for the Selective Identification and Quantification of the Amyloid Precursor Protein Tyr682 Residue Phosphorylation Status in Human Blood Mononuclear Cells |
| title_short |
Tandem Mass Spectrometry as Strategy for the Selective Identification and Quantification of the Amyloid Precursor Protein Tyr682 Residue Phosphorylation Status in Human Blood Mononuclear Cells |
| title_full |
Tandem Mass Spectrometry as Strategy for the Selective Identification and Quantification of the Amyloid Precursor Protein Tyr682 Residue Phosphorylation Status in Human Blood Mononuclear Cells |
| title_fullStr |
Tandem Mass Spectrometry as Strategy for the Selective Identification and Quantification of the Amyloid Precursor Protein Tyr682 Residue Phosphorylation Status in Human Blood Mononuclear Cells |
| title_full_unstemmed |
Tandem Mass Spectrometry as Strategy for the Selective Identification and Quantification of the Amyloid Precursor Protein Tyr682 Residue Phosphorylation Status in Human Blood Mononuclear Cells |
| title_sort |
tandem mass spectrometry as strategy for the selective identification and quantification of the amyloid precursor protein tyr682 residue phosphorylation status in human blood mononuclear cells |
| publisher |
MDPI AG |
| series |
Biomolecules |
| issn |
2218-273X |
| publishDate |
2021-08-01 |
| description |
<b>Background:</b> Alzheimer’s disease (AD) is a devastating neurodegenerative disease without guidelines for early diagnosis or personalized treatment. Previous studies have highlighted a crucial role of increasing phosphorylation levels of the amyloid precursor protein (APP) Tyr682 residue in predicting neuronal deficits in AD patients. However, the lack of a method for the identification and quantification of Tyr682 phosphorylation levels prevents its potential clinical applications. <b>Methods:</b> Here we report a method to identify and quantify APP Tyr682 phosphorylation levels in blood mononuclear cells of AD patients by tandem mass spectrometry (tMS). <b>Results:</b> This method showed excellent sensitivity with detection and quantification limits set respectively at 0.035 and 0.082 ng injected for the phosphorylated peptide and at 0.02 and 0.215 ng injected for the non-phosphorylated peptide. The average levels of both peptides were quantified in transfected HELA cells (2.48 and 3.53 ng/μg of protein, respectively). Preliminary data on 3 AD patients showed quantifiable levels of phosphorylated peptide (0.10–0.15 ng/μg of protein) and below the LOQ level of non-phosphorylated peptide (0.13 ng/μg of protein). <b>Conclusion:</b> This method could allow the identification of patients with increased APP Tyr682 phosphorylation and allow early characterization of molecular changes prior to the appearance of clinical signs. |
| topic |
Alzheimer’s disease APP Tyr682 phosphorylation targeted peptide analysis |
| url |
https://www.mdpi.com/2218-273X/11/9/1297 |
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