ArticleFASEB journal : official publication of the Federation of American Societies for Experimental Biology2012
A sensitive green fluorescent protein biomarker of N-glycosylation site occupancy.
Article in FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 27 citations in OpenAlex.
- Development of molecular sensors based on fluorescent proteins for polarized macrophages identification.Analytical sciences : the international journal of the Japan Society for Analytical Chemistry · 2024Article
- Molecular Spies in Action: Genetically Encoded Fluorescent Biosensors Light up Cellular Signals.Chemical reviews · 2024Review
- An RNA-centric dissection of host complexes controlling flavivirus infection.Nature microbiology · 2019Article
- CDG Therapies: From Bench to Bedside.International journal of molecular sciences · 2018Review
- Engineering posttranslational proofreading to discriminate nonstandard amino acids.Proceedings of the National Academy of Sciences of the United States of America · 2018Article
- Selenoprotein T is a novel OST subunit that regulates UPR signaling and hormone secretion.EMBO reports · 2017Article
- N-glycosylation Triggers a Dual Selection Pressure in Eukaryotic Secretory Proteins.Scientific reports · 2017Article
- N-Glycoprofiling Analysis for Carbohydrate Composition and Site-Occupancy Determination in a Poly-Glycosylated Protein: Human Thyrotropin of Different Origins.International journal of molecular sciences · 2017Article
- Assessment of Hepatocellular Carcinoma Metastasis Glycobiomarkers Using Advanced Quantitative N-glycoproteome Analysis.Frontiers in physiology · 2017Article
- Glycosylation and stem cells: Regulatory roles and application of iPSCs in the study of glycosylation-related disorders.BioEssays : news and reviews in molecular, cellular and developmental biology · 2016Review
- Glycomic Characterization of Induced Pluripotent Stem Cells Derived from a Patient Suffering from Phosphomannomutase 2 Congenital Disorder of Glycosylation (PMM2-CDG).Molecular & cellular proteomics : MCP · 2016Article
- A Novel N-Tetrasaccharide in Patients with Congenital Disorders of Glycosylation, Including Asparagine-Linked Glycosylation Protein 1, Phosphomannomutase 2, and Mannose Phosphate Isomerase Deficiencies.Clinical chemistry · 2016Article
- Development of isoform-specific sensors of polypeptide GalNAc-transferase activity.The Journal of biological chemistry · 2014Article
- A new congenital disorder of glycosylation caused by a mutation in SSR4, the signal sequence receptor 4 protein of the TRAP complex.Human molecular genetics · 2014Article
- Multiple phenotypes in phosphoglucomutase 1 deficiency.The New England journal of medicine · 2014Article
- A catalytically essential motif in external loop 5 of the bacterial oligosaccharyltransferase PglB.The Journal of biological chemistry · 2014Article
- Mutations in STT3A and STT3B cause two congenital disorders of glycosylation.Human molecular genetics · 2013Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
N-glycosylation mediates many biological functions. Genetic defects in the N-glycosylation pathway cause >35 inherited human disorders called congenital disorders of glycosylation (CDGs). As a result, some N-glycosylation sites are unoccupied. Serum transferrin is a diagnostic marker for these patients, but there are no corresponding cellular markers to assess glycosylation competence. Therefore, we engineered a green fluorescent protein (GFP) construct to measure N-glycosylation site occupancy. We designed an endoplasmic reticulum-retained GFP biomarker whose fluorescence is lost when it is N-glycosylated due to steric hindrance by the glycan. This marker is a highly sensitive indicator of N-glycosylation site occupancy. In CDG cells carrying the GFP construct, a 25% decrease of glycosylation efficiency induces a 5-fold increase in fluorescence, while cDNA complementation of the genetic defect results in a 5-fold decrease in fluorescence. This engineered GFP detects impaired N-glycosylation in multiple cell lines, including CHO cells, HeLa cells, normal and patient fibroblasts, induced pluripotent stem cells (iPSCs), and human embryonic stem cells (hESCs). This marker is a highly sensitive tool to study N-glycosylation site occupancy. It can be used to screen for compounds that reverse poor N-glycosylation site occupancy.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.