Precision Glycoproteomics Reveals Distinctive N-Glycosylation in Human Spermatozoa

Xin, Miaomiao; You, Shanshan; Xu, Yintai; Shi, Wenhao; Zhu, Bojing; Shen, Jiechen; Wu, Jingyu; Li, Cheng; Chen, Zexuan; Su, Yuanjie; Shi, Juanzi; Sun, Shisheng

doi:10.1016/j.mcpro.2022.100214

Cited by 21 publications

(20 citation statements)

References 49 publications

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“…Similarly, fewer numbers of fucoses were detected on galectin-3-binding protein in other tissues, such as four fucoses in human and mouse liver, , and three in human serum and mouse kidney (Figure G). Yet, both clusterin and galectin-3-binding protein have been modified by ten and seven fucoses in human spermatozoa, respectively . These data indicated that N- glycans on clusterin and galectin-3-binding protein might be tissue-specific, and heavy fucosylation might be a distinctive feature of semen.…”

Section: Resultsmentioning

confidence: 88%

“…Count: the number of heavily fucosylated glycoproteins involved in each specific biological process and reactome pathway/all heavily fucosylated glycoproteins identified in the present study. (F,G) Maximal fucoses per glycan identified on clusterin (F) and galectin-3-binding protein (G) in human seminal plasma currently and previously, including spermatozoa, liver, serum, and mouse tissues . F: fucose; SP: seminal plasma; SPZ: spermatozoa.…”

Section: Resultsmentioning

confidence: 99%

“…Intact glycopeptides were first separated into 96 fractions through a gradient of mobile phases A (2% ACN/5 mM NH 4 COOH, pH 10) and B (90% ACN/5 mM NH 4 COOH, pH 10) at a flow rate of 0.2 mL/min. A total of 120 min was used for gradient elution: 0–2% B for 7 min, 2–8% B for 35 min, 8–16% B for 35 min, 16–35% B for 10 min, 35–95% B for 5 min, 95–95% B for 28 min. Then, IGPs were integrated into 12 fractions according to previous studies and dried by vacuum centrifuging.…”

Section: Materials and Methodsmentioning

confidence: 99%

“…The desalted peptide mixtures eluted from C18 columns (in 50% ACN/0.1% TFA) were adjusted to a final solvent composition of 80% ACN/1%TFA and glycopeptides enriched by in house cotton columns. 27 Briefly, cotton in a 200 μL pipet tip was conditioned by 80% ACN/1% TFA, and samples loaded onto cotton columns were eventually eluted in 0.1% formic acid (FA) solution. The concentrations of enriched IGPs were estimated by Ultra Trace Ultraviolet Spectrophotometer (Denovix, USA).…”

Section: Glycopeptide Enrichmentmentioning

confidence: 99%

“…The mobile phase flow rate was 0.2 μL/ min and consisted of 0.1% FA in ddH 2 O (A) and 0.1% FA/ 80% ACN (B). A complete run of 240 min was set as follows: 27 3−9% B for 88 min, 9−31% B for 120 min, 31−40% B for 15 min, 40−99% B for 4 min, 99% B for 13 min. The settings for intact glycopeptide analysis were referred to a previous study: 29 the spray voltage (+) was set at 2.3 kV.…”

Section: Lc-ms/ms Analysismentioning

confidence: 99%

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Precision Structural Interpretation of Site-Specific N-Glycans in Seminal Plasma

Xin

You

et al. 2022

J. Proteome Res.

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N-Linked glycoproteins are rich in seminal plasma, playing various essential roles in supporting sperm function and the fertilization process. However, the detailed information on these glycoproteins, particularly site-specific glycan structures, is still limited. In this study, a precision site-specific N-glycoproteome map of human seminal plasma was established by employing the site-specific glycoproteomic approach and a recently developed glycan structure interpretation software, StrucGP. A total of 9567 unique glycopeptides identified in human seminal plasma were composed of 773 N-linked glycan structures and 1019 N-glycosites from 620 glycoproteins. These glycans were comprised of four types of core structures and 13 branch structures. The majority of identified glycoproteins functioned in response to stimulus and immunity. As we reported in human spermatozoa, heavy fucosylation (fucose residues ≥6 per glycan) was also detected on seminal plasma glycoproteins such as clusterin and galectin-3-binding protein, which were involved in the immune response of biological processes and reactome pathways. Comparison of site-specific glycans between seminal plasma and spermatozoa revealed more complicated glycan structures in seminal plasma than in spermatozoa, even on their shared glycoproteins. These present data will be greatly beneficial for the in-depth structural and functional study of glycosylation in the male reproduction system.

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Section: Resultsmentioning

confidence: 88%

Section: Resultsmentioning

confidence: 99%

Section: Materials and Methodsmentioning

confidence: 99%

Section: Glycopeptide Enrichmentmentioning

confidence: 99%

Section: Lc-ms/ms Analysismentioning

confidence: 99%

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Precision Structural Interpretation of Site-Specific N-Glycans in Seminal Plasma

Xin

You

et al. 2022

J. Proteome Res.

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Developing glycoproteomics reveals the role of posttranslational glycosylation in the physiological and pathological processes of male reproduction

Cheng,

Luo,

et al. 2024

iMetaOmics

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Glycosylation plays a pivotal role in the physiological and pathological processes of male reproduction. It impacts thousands of proteins and is actively ongoing through all stages of reproduction, including spermatogenesis, maturation, capacitation, and fertilization. However, our grasp on glycosylation within male reproductive processes remains limited, largely due to the technical hurdles. Recent advancements have seen the mapping of the glycoproteome of human semen, utilizing cutting‐edge glycoproteomic technologies. This breakthrough lays the groundwork for in‐depth research into the influence of glycosylation on male reproductive system and related disorders. Nevertheless, the field faces numerous challenges that necessitate further advancements in glycoproteomic methodologies. In this analysis, we evaluated the potential applications of advanced glycoproteomic techniques in the study of male reproduction and summarized the detailed profiling of the human semen glycome and glycoproteome. Our current understanding of glycosylation's role within the male reproductive system alongside recent progress in glycoproteomics may equip biologists with a comprehensive insight. Furthermore, this analysis brought together findings on abnormal glycosylation and its link to male reproductive disorders in the view of glycomics and glycoproteomics. It can facilitate the clinical application of glyco‐related biomarkers and targets in the treatment of infertility.

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Evaluation of absorbent cotton for glycopeptide enrichment

Xin

You

et al. 2022

Anal Bioanal Chem

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Precision Glycoproteomics Reveals Distinctive N-Glycosylation in Human Spermatozoa

Cited by 21 publications

References 49 publications

Precision Structural Interpretation of Site-Specific N-Glycans in Seminal Plasma

Precision Structural Interpretation of Site-Specific N-Glycans in Seminal Plasma

Developing glycoproteomics reveals the role of posttranslational glycosylation in the physiological and pathological processes of male reproduction

Evaluation of absorbent cotton for glycopeptide enrichment

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