The stem region of group A transferase is crucial for its specificity, and its alteration promotes heterologous Forssman synthase activity.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
26 08 2023
26 08 2023
Historique:
received:
15
02
2023
accepted:
17
08
2023
medline:
28
8
2023
pubmed:
27
8
2023
entrez:
26
8
2023
Statut:
epublish
Résumé
Some stem region mutants of human blood group A transferase (hAT) possess Forssman synthase (FS) activity, but very little is known about the mechanisms responsible for this enzymatic crosstalk. We performed confocal microscopy and image analysis to determine whether different intra-Golgi localization was accountable for this acquired activity. We also performed structural modeling and mutational and normal mode analyses. We introduced new mutations in the stem region and tested its FS and AT activities. No differences in subcellular localization were found between hAT and FS-positive mutants. AlphaFold models of hAT and mFS (mouse Forssman synthase) showed that the hAT stem region has a tether-like stem region, while in mFS, it encircles its catalytic domain. In silico analysis of FS-positive mutants indicated that stem region mutations induced structural changes, decreasing interatomic interactions and mobility of hAT that correlated with FS activity. Several additional mutations introduced in that region also bestowed FS activity without altering the AT activity: hAT 37-55 aa substitution by mFS 34-52, 37-55 aa deletion, and missense mutations: S46P, Q278Y, and Q286M. Stem region structure, mobility, and interactions are crucial for hAT specificity. Moreover, stem region mutations can lead to heterologous Forssman activity without changes in the catalytic machinery.
Identifiants
pubmed: 37634031
doi: 10.1038/s41598-023-40900-4
pii: 10.1038/s41598-023-40900-4
pmc: PMC10460411
doi:
Substances chimiques
globoside alpha-N-acetylgalactosaminyltransferase
EC 2.4.1.88
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
13996Informations de copyright
© 2023. Springer Nature Limited.
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