Structure of the human GlcNAc-1-phosphotransferase αβ subunits reveals regulatory mechanism for lysosomal enzyme glycan phosphorylation

Li, Hua; Lee, Wang-Sik; Feng, Xinwei; Bai, Lin; Jennings, Benjamin C.; Liu, Lin; Doray, Balraj; Canfield, William M.; Kornfeld, Stuart; H, Li

doi:10.1038/s41594-022-00748-0

Cited by 10 publications

(6 citation statements)

References 37 publications

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“…S4 C ). Additionally, during the review of this paper, the three-dimensional apo structure of human active GNPTAB, determined by cryogenic electron microscopy, was reported ( 68 ). Although the entire luminal portion was analyzed, only the catalytic core—approximately corresponding to the current crystal structure—was sufficiently ordered for high-resolution modeling ( SI Appendix , Fig.…”

Section: Resultsmentioning

confidence: 99%

Structures of the mannose-6-phosphate pathway enzyme, GlcNAc-1-phosphotransferase

Gorelik

Illés

Bui

et al. 2022

Proc. Natl. Acad. Sci. U.S.A.

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The mannose-6-phosphate (M6P) pathway is responsible for the transport of hydrolytic enzymes to lysosomes. N-acetylglucosamine-1-phosphotransferase (GNPT) catalyzes the first step of tagging these hydrolases with M6P, which when recognized by receptors in the Golgi diverts them to lysosomes. Genetic defects in the GNPT subunits, GNPTAB and GNPTG, cause the lysosomal storage diseases mucolipidosis types II and III. To better understand its function, we determined partial three-dimensional structures of the GNPT complex. The catalytic domain contains a deep cavity for binding of uridine diphosphate- N -acetylglucosamine, and the surrounding residues point to a one-step transfer mechanism. An isolated structure of the gamma subunit of GNPT reveals that it can bind to mannose-containing glycans in different configurations, suggesting that it may play a role in directing glycans into the active site. These findings may facilitate the development of therapies for lysosomal storage diseases.

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Section: Resultsmentioning

confidence: 99%

Structures of the mannose-6-phosphate pathway enzyme, GlcNAc-1-phosphotransferase

Gorelik

Illés

Bui

et al. 2022

Proc. Natl. Acad. Sci. U.S.A.

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“…This binding locks GNPT into an active state, initiating the phosphorylation reaction. Through this reaction, GNPT transfers the GlcNAc-1-phosphate residue from UDP-GlcNAc to the C-6-hydroxyl position of the terminal or penultimate mannose, forming a polysaccharide containing GlcNAc-P-Man ( Kudo et al., 2005 ; Kang et al., 2010 ; Gorelik et al., 2022 ; Li et al., 2022 ). Therefore, the knockout of GNPT results in the loss of M6P labeling and complete inactivation of lysosomal enzymes.…”

Section: The Components Of the M6p Pathwaymentioning

confidence: 99%

The host mannose-6-phosphate pathway and viral infection

Liu,

Wang,

et al. 2024

Front. Cell. Infect. Microbiol.

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Viruses, despite their simple structural composition, engage in intricate and complex interactions with their hosts due to their parasitic nature. A notable demonstration of viral behavior lies in their exploitation of lysosomes, specialized organelles responsible for the breakdown of biomolecules and clearance of foreign substances, to bolster their own replication. The man-nose-6-phosphate (M6P) pathway, crucial for facilitating the proper transport of hydrolases into lysosomes and promoting lysosome maturation, is frequently exploited for viral manipulation in support of replication. Recently, the discovery of lysosomal enzyme trafficking factor (LYSET) as a pivotal regulator within the lysosomal M6P pathway has introduced a fresh perspective on the intricate interplay between viral entry and host factors. This groundbreaking revelation illuminates unexplored dimensions of these interactions. In this review, we endeavor to provide a thorough overview of the M6P pathway and its intricate interplay with viral factors during infection. By consolidating the current understanding in this field, our objective is to establish a valuable reference for the development of antiviral drugs that selectively target the M6P pathway.

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“…How does TMEM251 deficiency lead to a general lysosome defect? We envisioned that two scenarios might explain this observation: (1) lysosomes might have an acidification defect, which inactivates lumenal hydrolases that depend on the low pH to be functional, or (2) the lumenal hydrolases might be absent from lysosomes. To test the first hypothesis, we stained cells with LysoTracker Red DND-99 that labeled the acidic endo-lysosomes and analyzed them by flow cytometry.…”

Section: Ablation Of Tmem251 Upregulates Lysosome Biogenesismentioning

confidence: 99%

“…It contains over 50 lumenal enzymes to carry out its hydrolysis function. Most enzymes utilize the mannose-6-phosphate (M6P) residues as a sorting signal for proper trafficking to the lysosome 1,2 .…”

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confidence: 99%

GCAF(TMEM251) regulates lysosome biogenesis by activating the mannose-6-phosphate pathway

Zhang

Yang

et al. 2022

Nat Commun

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The mannose-6-phosphate (M6P) biosynthetic pathway for lysosome biogenesis has been studied for decades and is considered a well-understood topic. However, whether this pathway is regulated remains an open question. In a genome-wide CRISPR/Cas9 knockout screen, we discover TMEM251 as the first regulator of the M6P modification. Deleting TMEM251 causes mistargeting of most lysosomal enzymes due to their loss of M6P modification and accumulation of numerous undigested materials. We further demonstrate that TMEM251 localizes to the Golgi and is required for the cleavage and activity of GNPT, the enzyme that catalyzes M6P modification. In zebrafish, TMEM251 deletion leads to severe developmental defects including heart edema and skeletal dysplasia, which phenocopies Mucolipidosis Type II. Our discovery provides a mechanism for the newly discovered human disease caused by TMEM251 mutations. We name TMEM251 as GNPTAB cleavage and activity factor (GCAF) and its related disease as Mucolipidosis Type V.

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Structure of the human GlcNAc-1-phosphotransferase αβ subunits reveals regulatory mechanism for lysosomal enzyme glycan phosphorylation

Cited by 10 publications

References 37 publications

Structures of the mannose-6-phosphate pathway enzyme, GlcNAc-1-phosphotransferase

Structures of the mannose-6-phosphate pathway enzyme, GlcNAc-1-phosphotransferase

The host mannose-6-phosphate pathway and viral infection

GCAF(TMEM251) regulates lysosome biogenesis by activating the mannose-6-phosphate pathway

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