Dataset Information

Analysis of intact mannose-6-phosphate glycopeptides

ABSTRACT: We have repurposed standard phosphoproteomics workflow based on Fe3+IMAC for enrichment ofmannose-6-phosphate modified glycopeptides. This worklfow was used to profile lysosomal acid hydrolases in HeLa and CHO cell lines. We have combined this approach with CRISPR/Cas9 KO of acid phosphatases 2 and 5 responsible for dephosphorylation of mannose-6-phosphate glycopeptides in the lysosome. This combined approach enabled significantly deeper coverage of CHO mannose-6-phosphate glycoproteome.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human) Cricetulus Griseus (chinese Hamster) (cricetulus Barabensis Griseus)

TISSUE(S): Cell Suspension Culture, Hela Cell

SUBMITTER: TOMISLAV CAVAL

LAB HEAD: Albert J R Heck

PROVIDER: PXD010333 | Pride | 2018-12-18

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20180409_462B2_01.raw_20180531_20ppm.mgf	Mgf
	20180409_462B2_01.raw_20180531_20ppm.mzid.gz	Mzid
	20180409_462B2_01.raw_20180531_20ppm.pride.mgf.gz	Mgf
	20180409_462B2_01.raw_20180531_20ppm.pride.mztab.gz	Mztab
	20180409_462B2_02.raw_20180531_20ppm.mgf	Mgf

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Publications

Targeted Analysis of Lysosomal Directed Proteins and Their Sites of Mannose-6-phosphate Modification.

Čaval Tomislav T Zhu Jing J Tian Weihua W Remmelzwaal Sanne S Yang Zhang Z Clausen Henrik H Heck Albert J R AJR

Molecular & cellular proteomics : MCP 20180920 1

Mannose-6-phosphate (M6P) is a distinctive post-translational modification critical for trafficking of lysosomal acid hydrolases into the lysosome. Improper trafficking into the lysosome, and/or lack of certain hydrolases, results in a toxic accumulation of their substrates within the lysosomes. To gain insight into the enzymes destined to the lysosome these glycoproteins can be distinctively enriched and studied using their unique M6P tag. Here we demonstrate, by adapting a protocol optimized f ...[more]

PMID: 30237200

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