Title : Quantitative comparison of PCa-associated glycan-com
positions and
lectin blotting analysis
Abstract :
- We compared the relative abundance of all intact N-glycopeptides grouped by glycan class and terminal features from PCa and BPH urine
- In doing this, no significant differences were observed between PCa and BPH with respect to the overall relative abundance of high mannose , afucosylated/asialylated complex/hybrid (C/H), fucosylated complex/hybrid (C/H\+Fuc) sialylated complex/hybrid (C/H\+NeuAc) and fucosylated/sialylated complex/hybrid (C/H\+Fuc/Sia)
- However, increased intensity of Fuc-containing intact desialo-O-glycopeptides was observed (T-test, p < 0.05) (Supplementary Figure 3)
- Comparative lectin blotting was also performed using concanavalin A (Con A), Maackia amurensis lectin ( MAL ), wheat germ agglutinin (WGA), Aleuria aurantia lectin (AAL) and Ricinus communis agglutinin (RCA) to assess the level of a range of glycoepitopes in PCa and BPH urine in an orthogonal manner
- After protein normalization, significantly increased levels of AAL reactivity were observed for the PCa urinary glycoproteome indicating a higher degree of core fucosylation (Figure 3)
- Slightly higher Con A reactivity was also observed in PCa relative to BPH urine, a feature not reflected in the quantitative LC-MS/MS data of high mannose glycopeptides
- However, Con A may also cross-react with paucimannosidic and biantennary complex glycoproteins possibly explaining this discrepancy [35]
- Based on these observations, we therefore sought to determine whether site-specific glycosylation instead would display a higher potential of discriminating between urine from PCa and BPH patients
Output (sent_index, trigger,
protein,
sugar,
site):
- 1. N-glycopeptides, , -, -, N-glycopeptides
- 2. fucosylated, , -, C/H+Fuc, -
- 2. fucosylated/sialylated, , -, C/H+Fuc/Sia, -
- 2. sialylated, , -, C/H+NeuAc, -
- 3. desialo-O-glycopeptides, , -, -, desialo-O-glycopeptides
- 6. glycopeptides, , -, -, glycopeptides
- 7. glycoproteins, , glycoproteins, -, -
Output(Part-Of) (sent_index,
protein,
site):
*Output_Site_Fusion* (sent_index,
protein,
sugar,
site):