pr9b00332_si_002.xlsx (56.44 kB)
Highly Efficient Analysis of Glycoprotein Sialylation in Human Serum by Simultaneous Quantification of Glycosites and Site-Specific Glycoforms
dataset
posted on 2019-08-14, 13:41 authored by Hongqiang Qin, Xuefang Dong, Jiawei Mao, Yao Chen, Mingming Dong, Liming Wang, Zhimou Guo, Xinmiao Liang, Mingliang YeAberrant sialylation
of glycoproteins is closely related to many
malignant diseases, and analysis of sialylation has great potential
to reveal the status of these diseases. However, in-depth analysis
of sialylation is still challenging because of the high microheterogeneity
of protein glycosylation, as well as the low abundance of sialylated
glycopeptides (SGPs). Herein, an integrated strategy was fabricated
for the detailed characterization of glycoprotein sialylation on the
levels of glycosites and site-specific glycoforms by employing the
SGP enrichment method. This strategy enabled the identification of
up to 380 glycosites, as well as 414 intact glycopeptides corresponding
to 383 site-specific glycoforms from only initial 6 μL serum
samples, indicating the high sensitivity of the method for the detailed
analysis of glycoprotein sialylation. This strategy was further employed
to the differential analysis of glycoprotein sialylation between hepatocellular
carcinoma patients and control samples, leading to the quantification
of 344 glycosites and 405 site-specific glycoforms, simultaneously.
Among these, 43 glycosites and 55 site-specific glycoforms were found
to have significant change on the glycosite and site-specific glycoform
levels, respectively. Interestingly, several glycoforms attached onto
the same glycosite were found with different change tendencies. This
strategy was demonstrated to be a powerful tool to reveal subtle differences
of the macro- and microheterogeneity of glycoprotein sialylation.