9f5h | pdb_00009f5h

Crystal structure of MGAT5 bump-and-hole mutant in complex with UDP and M592

spinqualitylabelsall models 9f5h, resolution 1.97Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 9f5h is a 2 chain structure with sequence from Homo sapiens. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 1.97Å Ligands: MAN, NAG, SO4, UDP Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function MGT5A_HUMAN Catalyzes the addition of N-acetylglucosamine in beta 1-6 linkage to the alpha-linked mannose of biantennary N-linked oligosaccharides. It is one of the most important enzymes involved in the regulation of the biosynthesis of glycoprotein oligosaccharides. Publication Abstract from PubMed Correct elaboration of N-linked glycans in the secretory pathway of human cells is essential in physiology. Early N-glycan biosynthesis follows an assembly line principle before undergoing crucial elaboration points that feature the sequential incorporation of the sugar N-acetylglucosamine (GlcNAc). The activity of GlcNAc transferase V (MGAT5) primes the biosynthesis of an N-glycan antenna that is heavily upregulated in cancer. Still, the functional relevance and substrate choice of MGAT5 are ill-defined. Here, we employ protein engineering to develop a bioorthogonal substrate analog for the activity of MGAT5. Chemoenzymatic synthesis is used to produce a collection of nucleotide-sugar analogs with bulky, bioorthogonal acylamide side chains. We find that WT-MGAT5 displays considerable activity toward such substrate analogues. Protein engineering yields an MGAT5 variant that loses activity against the native nucleotide sugar and increases activity toward a 4-azidobutyramide-containing substrate analogue. By such restriction of substrate specificity, we show that the orthogonal enzyme-substrate pair is suitable to bioorthogonally tag glycoproteins. Through X-ray crystallography and molecular dynamics simulations, we establish the structural basis of MGAT5 engineering, informing the design rules for bioorthogonal precision chemical tools. A Bioorthogonal Precision Tool for Human N-Acetylglucosaminyltransferase V.,Liu Y, Bineva-Todd G, Meek RW, Mazo L, Piniello B, Moroz O, Burnap SA, Begum N, Ohara A, Roustan C, Tomita S, Kjaer S, Polizzi K, Struwe WB, Rovira C, Davies GJ, Schumann B J Am Chem Soc. 2024 Oct 2;146(39):26707-26718. doi: 10.1021/jacs.4c05955. Epub , 2024 Sep 17. PMID:39287665 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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