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Optimized mucin-selective enrichment strategy to probe the mucinome

ABSTRACT: Mucin-domain glycoproteins are densely O-glycosylated and play critical roles in a host of biological functions. Previously, we developed a mucin-selective enrichment strategy by employing a catalytically inactive mucinase (StcE) conjugated to solid support. While this method was effective, it suffered from low throughput and high sample requirements. Further, we recently introduced a new mucinase (SmE) and demonstrated it has complementary recognition patterns to StcE. Here, we optimized our previous enrichment method to increase throughput and lower sample input amounts while maintaining mucin selectivity and enhancing glycopeptide signal. We also compare the ability of different mucinases, lectins, and mucin binding domains to enrich mucins from various cell lines and human serum. Overall, we demonstrate that our improved method StcE effectively isolates more mucin-domain glycoproteins, resulting in a high number of O-glycopeptides, which will allow for enhanced analysis of the mucinome.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Blood Serum

SUBMITTER: Vincent Chang

LAB HEAD: Vincent Chang

PROVIDER: PXD046534 | Pride | 2024-04-10

REPOSITORIES: Pride

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			Action	DRS
	210810_KEM_StcE_HeLa_HOAcNHS_Tryp_AC3PC2_Queen.raw	Raw
	210810_KEM_StcE_HeLa_UreaNHS_Tryp_AC3PC2_Queen.raw	Raw
	220121_KM_StcE_NHS_SDC_EDTA_SmE.raw	Raw
	230503_VC_StcEenrichserum_Trypsin.raw	Raw
	230503_VC_StcEenrichserum_Trypsin.raw_5_4_23_ StceEnrichTrypsin_mod(3).xlsx	Xlsx

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