The stem region of group A transferase is crucial for its specificity, and its alteration promotes heterologous Forssman synthase activity
Abstract 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 l...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-08-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-40900-4 |
| _version_ | 1797559879224786944 |
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| author | Emili Cid Miyako Yamamoto Laura Barrero Fumiichiro Yamamoto |
| author_facet | Emili Cid Miyako Yamamoto Laura Barrero Fumiichiro Yamamoto |
| author_sort | Emili Cid |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-03-10T17:51:29Z |
| format | Article |
| id | doaj.art-09afb5e27e7e435cb74354cdf0e0952d |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-10T17:51:29Z |
| publishDate | 2023-08-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-09afb5e27e7e435cb74354cdf0e0952d2023-11-20T09:19:38ZengNature PortfolioScientific Reports2045-23222023-08-0113111610.1038/s41598-023-40900-4The stem region of group A transferase is crucial for its specificity, and its alteration promotes heterologous Forssman synthase activityEmili Cid0Miyako Yamamoto1Laura Barrero2Fumiichiro Yamamoto3Laboratory of Immunohematology and Glycobiology, Josep Carreras Leukaemia Research Institute, Ctra. de Can RutiLaboratory of Immunohematology and Glycobiology, Josep Carreras Leukaemia Research Institute, Ctra. de Can RutiLaboratory of Immunohematology and Glycobiology, Josep Carreras Leukaemia Research Institute, Ctra. de Can RutiLaboratory of Immunohematology and Glycobiology, Josep Carreras Leukaemia Research Institute, Ctra. de Can RutiAbstract 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.https://doi.org/10.1038/s41598-023-40900-4 |
| spellingShingle | Emili Cid Miyako Yamamoto Laura Barrero Fumiichiro Yamamoto The stem region of group A transferase is crucial for its specificity, and its alteration promotes heterologous Forssman synthase activity Scientific Reports |
| title | The stem region of group A transferase is crucial for its specificity, and its alteration promotes heterologous Forssman synthase activity |
| title_full | The stem region of group A transferase is crucial for its specificity, and its alteration promotes heterologous Forssman synthase activity |
| title_fullStr | The stem region of group A transferase is crucial for its specificity, and its alteration promotes heterologous Forssman synthase activity |
| title_full_unstemmed | The stem region of group A transferase is crucial for its specificity, and its alteration promotes heterologous Forssman synthase activity |
| title_short | The stem region of group A transferase is crucial for its specificity, and its alteration promotes heterologous Forssman synthase activity |
| title_sort | stem region of group a transferase is crucial for its specificity and its alteration promotes heterologous forssman synthase activity |
| url | https://doi.org/10.1038/s41598-023-40900-4 |
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