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LC-MS/MS quantitation of esophagus disease blood serum glycoproteins by enrichment with hydrazide chemistry and lectin affinity chromatography.

Song E, Zhu R, Hammoud ZT, Mechref Y - J. Proteome Res. (2014)

Bottom Line: The quantitation and evaluation of significantly changed glycoproteins/glycopeptides are complementary between LAC and HC enrichments.LC-ESI-MS/MS analyses indicated that 7 glycoproteins enriched by LAC and 11 glycoproteins enriched by HC showed significantly different abundances between disease-free and disease cohorts.Multiple reaction monitoring quantitation resulted in 13 glycopeptides by LAC enrichment and 10 glycosylation sites by HC enrichment to be statistically different among disease cohorts.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry and Biochemistry, Texas Tech University , Memorial Circle & Boston, Lubbock, Texas 79409, United States.

ABSTRACT
Changes in glycosylation have been shown to have a profound correlation with development/malignancy in many cancer types. Currently, two major enrichment techniques have been widely applied in glycoproteomics, namely, lectin affinity chromatography (LAC)-based and hydrazide chemistry (HC)-based enrichments. Here we report the LC-MS/MS quantitative analyses of human blood serum glycoproteins and glycopeptides associated with esophageal diseases by LAC- and HC-based enrichment. The separate and complementary qualitative and quantitative data analyses of protein glycosylation were performed using both enrichment techniques. Chemometric and statistical evaluations, PCA plots, or ANOVA test, respectively, were employed to determine and confirm candidate cancer-associated glycoprotein/glycopeptide biomarkers. Out of 139, 59 common glycoproteins (42% overlap) were observed in both enrichment techniques. This overlap is very similar to previously published studies. The quantitation and evaluation of significantly changed glycoproteins/glycopeptides are complementary between LAC and HC enrichments. LC-ESI-MS/MS analyses indicated that 7 glycoproteins enriched by LAC and 11 glycoproteins enriched by HC showed significantly different abundances between disease-free and disease cohorts. Multiple reaction monitoring quantitation resulted in 13 glycopeptides by LAC enrichment and 10 glycosylation sites by HC enrichment to be statistically different among disease cohorts.

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Comparisons of MRM quantitation for 17 common glycosylationsitebetween LAC and HC enrichments in terms of ratio between DF versusHGD and DF versus EAC.
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fig3: Comparisons of MRM quantitation for 17 common glycosylationsitebetween LAC and HC enrichments in terms of ratio between DF versusHGD and DF versus EAC.

Mentions: 57 LAC-enriched glycopeptidesand 83 HC-enriched glycopeptides containing 85 glycosylation siteswere evaluated by MRM quantitation. The list of glycopeptides includingthe name of glycoprotein, peptide backbone sequences, m/z values, ppm, and quantitative values is summarizedin Supplementary Table 2 in the Supporting Information (LAC-enriched glycopeptides) and 3 (HC-enriched glycopeptides).The complementary MRM quantitation targeted a total of 70 glycoproteins(Supplementary Figure 6A in the Supporting Information) with 112 glycosylation sites (Supplementary Figure 6B in the Supporting Information). Seventeen glycosylationsites appeared to be common from LAC- and HC-enriched glycopeptidomicquantitation. Figure 3 illustrated the comparisonsof quantitative values (ratio) of the 17 glycosylation sites betweenDF versus HGD and DF versus EAC by LAC and HC enrichment techniques.Because the raw or normalized intensities are not comparative fromtwo enrichments, the comparison focuses on the ratio between diseasesubjects. Uniprot entry name was used instead of full name of glycoproteinsfor convenience. Student t test was performed using95% confidence interval to evaluate variation between LAC and HC quantitativevalues. As a result, a comparable trend of changes in disease cohortswas seen between LAC and HC enrichment techniques, except for a singleglycosylation site at N156 for FETUA (N156/FETUA). The change of N241/HPTin DF versus EAC by LAC and that of N494/KLKB1 in DF versus HGD byHC enrichment showed a large variation, which might be originatingfrom samples.


LC-MS/MS quantitation of esophagus disease blood serum glycoproteins by enrichment with hydrazide chemistry and lectin affinity chromatography.

Song E, Zhu R, Hammoud ZT, Mechref Y - J. Proteome Res. (2014)

Comparisons of MRM quantitation for 17 common glycosylationsitebetween LAC and HC enrichments in terms of ratio between DF versusHGD and DF versus EAC.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4227547&req=5

fig3: Comparisons of MRM quantitation for 17 common glycosylationsitebetween LAC and HC enrichments in terms of ratio between DF versusHGD and DF versus EAC.
Mentions: 57 LAC-enriched glycopeptidesand 83 HC-enriched glycopeptides containing 85 glycosylation siteswere evaluated by MRM quantitation. The list of glycopeptides includingthe name of glycoprotein, peptide backbone sequences, m/z values, ppm, and quantitative values is summarizedin Supplementary Table 2 in the Supporting Information (LAC-enriched glycopeptides) and 3 (HC-enriched glycopeptides).The complementary MRM quantitation targeted a total of 70 glycoproteins(Supplementary Figure 6A in the Supporting Information) with 112 glycosylation sites (Supplementary Figure 6B in the Supporting Information). Seventeen glycosylationsites appeared to be common from LAC- and HC-enriched glycopeptidomicquantitation. Figure 3 illustrated the comparisonsof quantitative values (ratio) of the 17 glycosylation sites betweenDF versus HGD and DF versus EAC by LAC and HC enrichment techniques.Because the raw or normalized intensities are not comparative fromtwo enrichments, the comparison focuses on the ratio between diseasesubjects. Uniprot entry name was used instead of full name of glycoproteinsfor convenience. Student t test was performed using95% confidence interval to evaluate variation between LAC and HC quantitativevalues. As a result, a comparable trend of changes in disease cohortswas seen between LAC and HC enrichment techniques, except for a singleglycosylation site at N156 for FETUA (N156/FETUA). The change of N241/HPTin DF versus EAC by LAC and that of N494/KLKB1 in DF versus HGD byHC enrichment showed a large variation, which might be originatingfrom samples.

Bottom Line: The quantitation and evaluation of significantly changed glycoproteins/glycopeptides are complementary between LAC and HC enrichments.LC-ESI-MS/MS analyses indicated that 7 glycoproteins enriched by LAC and 11 glycoproteins enriched by HC showed significantly different abundances between disease-free and disease cohorts.Multiple reaction monitoring quantitation resulted in 13 glycopeptides by LAC enrichment and 10 glycosylation sites by HC enrichment to be statistically different among disease cohorts.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry and Biochemistry, Texas Tech University , Memorial Circle & Boston, Lubbock, Texas 79409, United States.

ABSTRACT
Changes in glycosylation have been shown to have a profound correlation with development/malignancy in many cancer types. Currently, two major enrichment techniques have been widely applied in glycoproteomics, namely, lectin affinity chromatography (LAC)-based and hydrazide chemistry (HC)-based enrichments. Here we report the LC-MS/MS quantitative analyses of human blood serum glycoproteins and glycopeptides associated with esophageal diseases by LAC- and HC-based enrichment. The separate and complementary qualitative and quantitative data analyses of protein glycosylation were performed using both enrichment techniques. Chemometric and statistical evaluations, PCA plots, or ANOVA test, respectively, were employed to determine and confirm candidate cancer-associated glycoprotein/glycopeptide biomarkers. Out of 139, 59 common glycoproteins (42% overlap) were observed in both enrichment techniques. This overlap is very similar to previously published studies. The quantitation and evaluation of significantly changed glycoproteins/glycopeptides are complementary between LAC and HC enrichments. LC-ESI-MS/MS analyses indicated that 7 glycoproteins enriched by LAC and 11 glycoproteins enriched by HC showed significantly different abundances between disease-free and disease cohorts. Multiple reaction monitoring quantitation resulted in 13 glycopeptides by LAC enrichment and 10 glycosylation sites by HC enrichment to be statistically different among disease cohorts.

Show MeSH
Related in: MedlinePlus