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| Pàgina inicial > Articles > Articles publicats > The stem region of group A transferase is crucial for its specificity, and its alteration promotes heterologous Forssman synthase activity |
(Institut Germans Trias i Pujol. Institut de Recerca contra la Leucèmia Josep Carreras) (Institut Germans Trias i Pujol. Institut de Recerca contra la Leucèmia Josep Carreras) (Institut Germans Trias i Pujol. Institut de Recerca contra la Leucèmia Josep Carreras)| Data: | 2023 |
| Resum: | 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. |
| Ajuts: | Agència de Gestió d'Ajuts Universitaris i de Recerca 2014 SGR 1269 Agència de Gestió d'Ajuts Universitaris i de Recerca 2017 SGR 529 Agència de Gestió d'Ajuts Universitaris i de Recerca 2021 SGR 00560 Instituto de Salud Carlos III PI11/00454 |
| Nota: | We thank Alba Garcia and Albert Pérez Ladaga for their technical assistance, and José Luis Gelpí Buchaca for reviewing the article. In addition, we thank Marco Antonio Fernández Sanmartín and Gerard Requena Fernández at the IGTP Cytometry Core Facility for their help with cytometry analyses and Jakub Chojnacki of the IGTP Microscopy Core facility for his assistance with the confocal imaging. The study was supported by the Spanish Health Research Foundation (Instituto de Salud Carlos III) grant (PI11/00454), the Departament de Recerca i Universitats of Generalitat de Catalunya through the fund from Agència de Gestió d'Ajuts Universitaris i de Recerca (2014 SGR 1269, 2017 SGR 529 and 2021 SGR 00560), and the institutional start-up funds from IMPPC and IJC-"La Caixa" Foundation to F.Y. IJC and IGTP are CERCA centers supported by CERCA Programme/Generalitat de Catalunya. Josep Carreras Foundation also supports IJC. |
| Drets: | Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.  |
| Llengua: | Anglès |
| Document: | Article ; recerca ; Versió publicada |
| Matèria: | Animals ; Catalysis ; Catalytic Domain ; Cross Reactions ; Humans ; Mice ; Mutation |
| Publicat a: | Scientific reports, Vol. 13 Núm. 1 (december 2023) , p. 13996, ISSN 2045-2322 |
