The luciferase-based in vivo protein–protein interaction assay revealed that CHK1 promotes PP2A and PME-1 interaction

Ando, Sana; Tanaka, Keiko; Matsumoto, Maharu; Oyama, Yuki; Tomabechi, Yuri; Yamagata, Atsushi; Shirouzu, Mikako; Nakagawa, Reiko; Okimoto, Noriaki; Taiji, Makoto; Sato, Koichi; Ohama, Takashi

doi:10.1016/j.jbc.2024.107277

Cited by 2 publications

(1 citation statement)

References 55 publications

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“…NanoBiT technology was used to assess the formation of homo-and heterodimers by high-frequency and mutein human A4galt. Although it is a relatively new method, NanoBiT has already been used to identify interaction partners for other GTs, such as human B4galt4 [31] and B4galt1 [32], as well as other proteins [33][34]. In our study, we used two cell lines, CHO-Lec2 and HEK293T.…”

Section: Analysis Of Human A4galt Dimerization Using Nanobit Technologymentioning

confidence: 99%

Delving into human α1,4-galactosyltransferase acceptor specificity: the role of enzyme dimerization

Mikołajczyk,

Wróblewski,

Kmiecik

2024

Preprint

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Glycosyltransferases (GTs) exhibit precise donor and acceptor specificities, governed by intricate mechanisms, including protein assembly. Human α1,4-galactosyltransferase (A4galt), a Golgi apparatus-resident GT, catalyzes the synthesis of Gb3 glycosphingolipid (GSL) and P1 glycotope on glycoproteins (GPs), receptors for Shiga toxin type 1 (Stx1) and 2 (Stx2). These toxins are produced by enterohemorrhagic Escherichia coli and can lead to severe outcomes, such as hemolytic-uremic syndrome (HUS). To elucidate the molecular basis of A4galt specificity towards GSLs and GPs, we explored its interaction with GP-specific β1,4-galactosyltransferase 1 (B4galt1) and GSL-specific β1,4-galactosyltransferase isoenzymes 5 and 6 (B4galt5 and B4galt6). Using a novel NanoBiT assay, we demonstrated that A4galt could form homodimers and heterodimers with B4galt1 and B4galt5. Furthermore, by employing AlphaFold for state-of-the-art structural prediction, we analyzed the interactions and structures of these enzyme complexes. Our analysis highlighted that the A4galt-B4galt5 heterodimer exhibited the highest prediction confidence, indicating a significant role of A4galt heterodimers in determining enzyme specificity toward GSLs and GPs. These insights not only increase our knowledge about glycosylation processes but also open avenues for developing new therapeutic approaches to Shiga toxin-related diseases.

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Section: Analysis Of Human A4galt Dimerization Using Nanobit Technologymentioning

confidence: 99%

Delving into human α1,4-galactosyltransferase acceptor specificity: the role of enzyme dimerization

Mikołajczyk,

Wróblewski,

Kmiecik

2024

Preprint

View full text Add to dashboard Cite

show abstract

Delving into human α1,4-galactosyltransferase acceptor specificity: The role of enzyme dimerization

Mikołajczyk,

Wróblewski,

Kmiecik

2024

Biochemical and Biophysical Research Communications

View full text Add to dashboard Cite

The luciferase-based in vivo protein–protein interaction assay revealed that CHK1 promotes PP2A and PME-1 interaction

Cited by 2 publications

References 55 publications

Delving into human α1,4-galactosyltransferase acceptor specificity: the role of enzyme dimerization

Delving into human α1,4-galactosyltransferase acceptor specificity: the role of enzyme dimerization

Delving into human α1,4-galactosyltransferase acceptor specificity: The role of enzyme dimerization

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