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Determination of GPI structure of human CD59 in HEK293 cells

ABSTRACT: To determine GPI structure of human CD59 in wild-type,B3GALT4-KO, PIGZ-B3GALT4-DKO HEK293 cells.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell, Epithelial Cell

SUBMITTER: Yicheng Wang

LAB HEAD: Taroh Kinoshita

PROVIDER: PXD014226 | Pride | 2020-02-17

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	170706_KO.dat	Other
	170706_KO.mgf	Mgf
	170706_KO.raw	Raw
	170706_WT.dat	Other
	170706_WT.mgf	Mgf

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1 - 5 of 18

Publications

Cross-talks of glycosylphosphatidylinositol biosynthesis with glycosphingolipid biosynthesis and ER-associated degradation.

Wang Yicheng Y Maeda Yusuke Y Liu Yi-Shi YS Takada Yoko Y Ninomiya Akinori A Hirata Tetsuya T Fujita Morihisa M Murakami Yoshiko Y Kinoshita Taroh T

Nature communications 20200213 1

Glycosylphosphatidylinositol (GPI)-anchored proteins and glycosphingolipids interact with each other in the mammalian plasma membranes, forming dynamic microdomains. How their interaction starts in the cells has been unclear. Here, based on a genome-wide CRISPR-Cas9 genetic screen for genes required for GPI side-chain modification by galactose in the Golgi apparatus, we report that β1,3-galactosyltransferase 4 (B3GALT4), the previously characterized GM1 ganglioside synthase, additionally functio ...[more]

PMID: 32054864

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Determination of GPI structure of human CD59 in various KO CHO cells

Project description:To determine GPI structure of human CD59 in PGAP4-KO, PGAP3-KO, PGAP5-KO, and PGAP2-KO CHO cells.

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A novel structure of GPI anchored proteins

Project description:GPI anchors many proteins to the cell surface. GPI precursor has three mannoses, all of which are modified by ethanolamine-phosphate (EthN-P). It has been believed that EthN-P on the third mannose is always used as a bridge to the protein and EthN-P on the second mannose is removed after GPI is attached to the protein. Based on the finding that PIGB-knockout cells lacking the third mannose and PIGO-knockout cells lacking EthN-P linked to the third mannose expressed low levels of CD59 and DAF, GPI-APs and that those residual GPI-APs were lost by further knockout of PIGG. Using mass-spectrometry we determined that CD59 expressed on PIGB-knockout cells is attached to GPI via EthN-P linked to the second mannose. This linkage structure is also found in wild type cells. Our data modifies the current view of GPI anchors and provides mechanistic basis of inherited GPI deficiency caused by PIGG mutations.

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GPI-glycan structures of Prion from PGAP4-KO mouse brains

Project description:To determine glycosylphosphatidylinositol (GPI) structures of prion proteins isolated from PGAP4-KO mouse brains.

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GPI-glycan structures of Prion from WT mouse brains

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GPI-glycan structures of human Prion (M232R) from knock-in mice_LC-MSMS

Project description:To determine glycosylphosphatidylinositol (GPI) structures of human prion proteins with M232R mutation isolated from knock-in mice those express human prion protein instead of mouse one.

2023-03-08 | PXD037584 | Pride

GPI-glycan structures of human Prion from a human brain

Project description:To determine glycosylphosphatidylinositol (GPI) structures of human prion proteins isolated from a human brain.

2020-04-22 | PXD017656 | Pride

GPI-glycan structures of human Prion from knock-in mice_LC-MSMS

Project description:To determine glycosylphosphatidylinositol (GPI) structures of human prion proteins isolated from knock-in mice those express human prion protein instead of mouse one.

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A novel structure of GPI anchored proteins-NT5E

Project description:GPI anchors many proteins to the cell surface. GPI precursor has three mannoses, all of which are modified by ethanolamine-phosphate (EthN-P). It has been believed that EthN-P on the third mannose is always used as a bridge to the protein and EthN-P on the second mannose is removed after GPI is attached to the protein. In fact, several GPI-anchored proteins are not appreciably reduced on cells defective in PIGG, which transfers EthN-P to the second mannose. Nevertheless, mutations in PIGG cause neuronal abnormalities. Here, we show that EthN-P on the second mannose is used as a preferential bridge for several GPI-anchored proteins. Our data modifies the current view of GPI anchors and provides mechanistic basis of PIGG deficiencies.

2022-05-31 | PXD029850 | Pride

A novel structure of GPI anchored proteins NTNG2

Project description:GPI anchors many proteins to the cell surface. GPI precursor has three mannoses, all of which are modified by ethanolamine-phosphate (EthN-P). It has been believed that EthN-P on the third mannose is always used as a bridge to the protein and EthN-P on the second mannose is removed after GPI is attached to the protein. In fact, several GPI-anchored proteins are not appreciably reduced on cells defective in PIGG, which transfers EthN-P to the second mannose. Nevertheless, mutations in PIGG cause neuronal abnormalities. Here, we show that EthN-P on the second mannose is used as a preferential bridge for several GPI-anchored proteins. Our data modifies the current view of GPI anchors and provides mechanistic basis of PIGG deficiencies.

2022-05-31 | PXD029851 | Pride

Expression microarray analysis of human pancreatic islets reveals CD59 function

Project description:Pancreatic islets are central in type 2-diabetes development, which coincides with increased activity of innate immunity. Intriguingly, human pancreatic islets express many complement genes. The most highly expressed gene was the complement inhibitor CD59 that is GPI anchored to the cell membrane, which unexpectedly was found in high amounts intracellularly in beta cells. Silencing of CD59 strongly suppressed insulin secretion. Importantly, this suppression was unrelated to established CD59 functions, but rather depletion of intracellular CD59. Imaging experiments identified a distal site of inhibition in the exocytotic pathway, but prior to emptying of the insulin granules. Proximity Ligation Assays pin-pointed the mechanism to impaired turnover of exocytosis-regulating SNARE-proteins and CD59 was detected in complex with VAMP2 and syntaxin. CD59 was downregulated by 24-h glucose incubations in human islets, rat cell lines and in islets from three rodent diabetes models. Islets from cadaver donors were provided by the Nordic Islet Transplantation Programme (www.nordicislets.org), Uppsala University. The microarrays were performed using GeneChipM-BM-. Human Gene 1.0 ST whole transcript according to Affymetrix standard protocol.

2014-03-02 | E-GEOD-54279 | biostudies-arrayexpress

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