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The tryptic peptides of hemoglobin for diagnosis of type 2 diabetes mellitus using label-free and standard-free LC-ESI-DMRM

N-1-(deoxyfructosyl) valine of β-hemoglobin, commonly referred to as HbA1c, is the “gold standard” for clinical detection of diabetes. Instead of quantifying the full-length HbA1c glycated protein, in the present study, we proposed the peptide-based strategy to quantify the depletion of the tryptic...

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Autores principales: Hong, Jie, Xin, Shuchen, Min, Rui, Zhang, Yongqian, Deng, Yulin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8102995/
https://www.ncbi.nlm.nih.gov/pubmed/33932868
http://dx.doi.org/10.1016/j.redox.2021.101985
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author Hong, Jie
Xin, Shuchen
Min, Rui
Zhang, Yongqian
Deng, Yulin
author_facet Hong, Jie
Xin, Shuchen
Min, Rui
Zhang, Yongqian
Deng, Yulin
author_sort Hong, Jie
collection PubMed
description N-1-(deoxyfructosyl) valine of β-hemoglobin, commonly referred to as HbA1c, is the “gold standard” for clinical detection of diabetes. Instead of quantifying the full-length HbA1c glycated protein, in the present study, we proposed the peptide-based strategy to quantify the depletion of the tryptic peptides of hemoglobin for the diagnosis of type 2 diabetes mellitus (T2DM). The peptides were discovered and validated as T2DM biomarkers by label-free LC-ESI-DMRM method without reference material. The glucose could react with hemoglobin's free amino group of N-terminus and ϵ-amino group of lysine residues and leave the modification on the hemoglobin tryptic peptides. Thus, there are two types of peptides in the hemoglobin: sensitive peptides and insensitive peptides to glucose due to the differential sensitivity of lysine residues to glycation. To discover two types of peptides of hemoglobin, we first developed the assay of liquid chromatography-electrospray ionization mass spectrometry coupled with dynamic multiple reaction monitoring. The protein coverage reaches 94.2%. Moreover, the hemoglobin was incubated with the 500 mmol/L glucose for 20 days, 40 days and 60 days in vitro to screen the sensitive peptides and insensitive peptides to glucose. A total of 14 sensitive peptides and 4 insensitive peptides were discovered. Furthermore, the LC-ESI-DMRM method was also utilized to validated the glucose-sensitive peptides by 40 clinical samples with healthy control individuals (n = 20) and type 2 diabetes mellitus patients (n = 20). Three putative sensitive peptides (LLGNVLVCVLAHHFGK, VVAGVANALAHKYH, LRVDPVNFK) from the hemoglobin showed excellent sensitivity and specificity based on receiver operating characteristic analysis and were verified as the promising biomarkers for the diagnosis of diabetes mellitus. And one peptide (LLVVYPWTQR) was found as glucose-insensitive peptide. Taken together, the findings of this study suggest that quantification of hemoglobin tryptic peptides using label-free and standard-free LC-ESI-DMRM is an alternative method for the diagnosis of T2DM, which could be combined with other MS-based blood biomarkers for diagnosis of multiple diseases in MS single shot.
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spelling pubmed-81029952021-05-14 The tryptic peptides of hemoglobin for diagnosis of type 2 diabetes mellitus using label-free and standard-free LC-ESI-DMRM Hong, Jie Xin, Shuchen Min, Rui Zhang, Yongqian Deng, Yulin Redox Biol Research Paper N-1-(deoxyfructosyl) valine of β-hemoglobin, commonly referred to as HbA1c, is the “gold standard” for clinical detection of diabetes. Instead of quantifying the full-length HbA1c glycated protein, in the present study, we proposed the peptide-based strategy to quantify the depletion of the tryptic peptides of hemoglobin for the diagnosis of type 2 diabetes mellitus (T2DM). The peptides were discovered and validated as T2DM biomarkers by label-free LC-ESI-DMRM method without reference material. The glucose could react with hemoglobin's free amino group of N-terminus and ϵ-amino group of lysine residues and leave the modification on the hemoglobin tryptic peptides. Thus, there are two types of peptides in the hemoglobin: sensitive peptides and insensitive peptides to glucose due to the differential sensitivity of lysine residues to glycation. To discover two types of peptides of hemoglobin, we first developed the assay of liquid chromatography-electrospray ionization mass spectrometry coupled with dynamic multiple reaction monitoring. The protein coverage reaches 94.2%. Moreover, the hemoglobin was incubated with the 500 mmol/L glucose for 20 days, 40 days and 60 days in vitro to screen the sensitive peptides and insensitive peptides to glucose. A total of 14 sensitive peptides and 4 insensitive peptides were discovered. Furthermore, the LC-ESI-DMRM method was also utilized to validated the glucose-sensitive peptides by 40 clinical samples with healthy control individuals (n = 20) and type 2 diabetes mellitus patients (n = 20). Three putative sensitive peptides (LLGNVLVCVLAHHFGK, VVAGVANALAHKYH, LRVDPVNFK) from the hemoglobin showed excellent sensitivity and specificity based on receiver operating characteristic analysis and were verified as the promising biomarkers for the diagnosis of diabetes mellitus. And one peptide (LLVVYPWTQR) was found as glucose-insensitive peptide. Taken together, the findings of this study suggest that quantification of hemoglobin tryptic peptides using label-free and standard-free LC-ESI-DMRM is an alternative method for the diagnosis of T2DM, which could be combined with other MS-based blood biomarkers for diagnosis of multiple diseases in MS single shot. Elsevier 2021-04-22 /pmc/articles/PMC8102995/ /pubmed/33932868 http://dx.doi.org/10.1016/j.redox.2021.101985 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Paper
Hong, Jie
Xin, Shuchen
Min, Rui
Zhang, Yongqian
Deng, Yulin
The tryptic peptides of hemoglobin for diagnosis of type 2 diabetes mellitus using label-free and standard-free LC-ESI-DMRM
title The tryptic peptides of hemoglobin for diagnosis of type 2 diabetes mellitus using label-free and standard-free LC-ESI-DMRM
title_full The tryptic peptides of hemoglobin for diagnosis of type 2 diabetes mellitus using label-free and standard-free LC-ESI-DMRM
title_fullStr The tryptic peptides of hemoglobin for diagnosis of type 2 diabetes mellitus using label-free and standard-free LC-ESI-DMRM
title_full_unstemmed The tryptic peptides of hemoglobin for diagnosis of type 2 diabetes mellitus using label-free and standard-free LC-ESI-DMRM
title_short The tryptic peptides of hemoglobin for diagnosis of type 2 diabetes mellitus using label-free and standard-free LC-ESI-DMRM
title_sort tryptic peptides of hemoglobin for diagnosis of type 2 diabetes mellitus using label-free and standard-free lc-esi-dmrm
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8102995/
https://www.ncbi.nlm.nih.gov/pubmed/33932868
http://dx.doi.org/10.1016/j.redox.2021.101985
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