Prediction and verification of glycosyltransferase activity by bioinformatics analysis and protein engineering

Gerloff, Dietlind L.; Ilina, Elena I.; Cialini, Camille; Salcedo, Uxue Mata; Mittelbronn, Michel; Müller, Tanja

doi:10.1016/j.xpro.2022.101905

Cited by 1 publication

(2 citation statements)

References 35 publications

(96 reference statements)

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“…Prior efforts at modeling the GLT8D1 structure revealed a GT-A fold for the enzyme 49 , including conserved landmark features 15 , and the absence of an xED motif sequence that harbors the catalytic base in inverting enzymes that is consistent with the retaining mechanism for the GT8 family enzymes 22 . We employed an AlphaFold2 model of GLT8D1 to query structure databases and identify well-characterized enzymes with high similarity to GLT8D1.…”

Section: Discussionmentioning

confidence: 99%

“…In view of the several open possibilities, it is necessary to investigate further to find efficient endogenous GLT8D1 substrates, to structurally characterize them and evaluate their role in the mechanisms underlying diseases of the central nervous system and cancer. Prior efforts at modeling the GLT8D1 structure revealed a GT-A fold for the enzyme 49 , including conserved landmark features 15 , and the absence of an xED motif sequence that harbors the catalytic base in inverting enzymes that is consistent with the retaining mechanism for the GT8 family enzymes 22 . We employed an AlphaFold2 model of GLT8D1 to query structure databases and identify well-characterized enzymes with high similarity to GLT8D1.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Glycosyltransferase 8 domain-containing protein 1 (GLT8D1) is a UDP-dependent galactosyltransferase

Vicente,

Guerreiro,

Felgueiras

et al. 2023

Sci Rep

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Glycosyltransferases (GTs) are enzymes that catalyze the formation of glycosidic bonds and hundreds of GTs have been identified so far in humans. Glycosyltransferase 8 domain-containing protein 1 (GLT8D1) has been associated with central nervous system diseases and cancer. However, evidence on its enzymatic properties, including its substrates, has been scarcely described. In this paper, we have produced and purified recombinant secretory GLT8D1. The enzyme was found to be N-glycosylated. Differential scanning fluorimetry was employed to analyze the stabilization of GLT8D1 by Mn2+ and nucleotides, revealing UDP as the most stabilizing nucleotide scaffold. GLT8D1 displayed glycosyltransferase activity from UDP-galactose onto N-acetylgalactosamine but with a low efficiency. Modeling of the structure revealed similarities with other GT-A fold enzymes in CAZy family GT8 and glycosyltransferases in other families with galactosyl-, glucosyl-, and xylosyltransferase activities, each with retaining catalytic mechanisms. Our study provides novel structural and functional insights into the properties of GLT8D1 with implications in pathological processes.

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Section: Discussionmentioning

confidence: 99%