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Proximal partners of the organellar N-terminal acetyltransferase NAA60 : insights into Golgi structure and transmembrane protein topology

Sebastian Tanco (UGent) , Veronique Jonckheere (UGent) , Arun Kumar Tharkeshwar Raghunath, Annelies Bogaert, Kris Gevaert (UGent) , Wim Anneart and Petra Van Damme (UGent)
(2025) OPEN BIOLOGY. 15(2).
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Abstract
Biotin identification (BioID) is an interactomics approach that utilizes proximity labelling to map the local interactome or proxeome of proteins within a cell. This study applies BioID to investigate proteins proximal to NAA60 (N-alpha-acetyltransferase 60), an N-terminal acetyltransferase (NAT) of pathological significance in human disease, characterized by its unique Golgi localization. NAA60 is known to N-terminally acetylate transmembrane proteins that present their N-terminus on the cytosolic face of the membrane, and its involvement in maintaining Golgi structure has previously been established. Using a stable cell-line expressing an NAA60-BirA* fusion protein, we isolated biotinylated proteins through streptavidin affinity purification. Mass spectrometry analysis revealed over 100 proximal partners of NAA60, enriched in proteins localized on the trans-side of the Golgi apparatus. High-confidence proximity interactors included golgins and GRASP proteins, essential for Golgi integrity. Considering the transmembrane nature of NAA60, the identification of biotinylated peptides inferred the topology of transmembrane protein interactors within the secretory pathway. Subsequent suborganellar localization analysis revealed a more prominent medial/trans-Golgi localization of NAA60. Our findings underscore the role of NAA60 and its interactors in maintaining Golgi structural integrity and highlight the effectiveness of BioID in generating critical protein topology data, invaluable for enhancing the prediction of protein topology within cellular compartments.
Keywords
proximity-dependent biotin identification (BioID), Golgi fragmentation, NAA60, N-terminal acetylation, proxeome, transmembrane protein topology, ARF-LIKE GTPASE, ALPHA-ACETYLTRANSFERASE, MOLECULAR DETERMINANTS, ACETYLATION, IDENTIFICATION, TRAFFICKING, PLATFORM

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Citation

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MLA
Tanco, Sebastian, et al. “Proximal Partners of the Organellar N-Terminal Acetyltransferase NAA60 : Insights into Golgi Structure and Transmembrane Protein Topology.” OPEN BIOLOGY, vol. 15, no. 2, 2025, doi:10.1098/rsob.240225.
APA
Tanco, S., Jonckheere, V., Tharkeshwar Raghunath, A. K., Bogaert, A., Gevaert, K., Anneart, W., & Van Damme, P. (2025). Proximal partners of the organellar N-terminal acetyltransferase NAA60 : insights into Golgi structure and transmembrane protein topology. OPEN BIOLOGY, 15(2). https://doi.org/10.1098/rsob.240225
Chicago author-date
Tanco, Sebastian, Veronique Jonckheere, Arun Kumar Tharkeshwar Raghunath, Annelies Bogaert, Kris Gevaert, Wim Anneart, and Petra Van Damme. 2025. “Proximal Partners of the Organellar N-Terminal Acetyltransferase NAA60 : Insights into Golgi Structure and Transmembrane Protein Topology.” OPEN BIOLOGY 15 (2). https://doi.org/10.1098/rsob.240225.
Chicago author-date (all authors)
Tanco, Sebastian, Veronique Jonckheere, Arun Kumar Tharkeshwar Raghunath, Annelies Bogaert, Kris Gevaert, Wim Anneart, and Petra Van Damme. 2025. “Proximal Partners of the Organellar N-Terminal Acetyltransferase NAA60 : Insights into Golgi Structure and Transmembrane Protein Topology.” OPEN BIOLOGY 15 (2). doi:10.1098/rsob.240225.
Vancouver
1.
Tanco S, Jonckheere V, Tharkeshwar Raghunath AK, Bogaert A, Gevaert K, Anneart W, et al. Proximal partners of the organellar N-terminal acetyltransferase NAA60 : insights into Golgi structure and transmembrane protein topology. OPEN BIOLOGY. 2025;15(2).
IEEE
[1]
S. Tanco et al., “Proximal partners of the organellar N-terminal acetyltransferase NAA60 : insights into Golgi structure and transmembrane protein topology,” OPEN BIOLOGY, vol. 15, no. 2, 2025.
@article{01JPAK3E3EKJJVPPZRS544HK5D,
  abstract     = {{Biotin identification (BioID) is an interactomics approach that utilizes proximity labelling to map the local interactome or proxeome of proteins within a cell. This study applies BioID to investigate proteins proximal to NAA60 (N-alpha-acetyltransferase 60), an N-terminal acetyltransferase (NAT) of pathological significance in human disease, characterized by its unique Golgi localization. NAA60 is known to N-terminally acetylate transmembrane proteins that present their N-terminus on the cytosolic face of the membrane, and its involvement in maintaining Golgi structure has previously been established. Using a stable cell-line expressing an NAA60-BirA* fusion protein, we isolated biotinylated proteins through streptavidin affinity purification. Mass spectrometry analysis revealed over 100 proximal partners of NAA60, enriched in proteins localized on the trans-side of the Golgi apparatus. High-confidence proximity interactors included golgins and GRASP proteins, essential for Golgi integrity. Considering the transmembrane nature of NAA60, the identification of biotinylated peptides inferred the topology of transmembrane protein interactors within the secretory pathway. Subsequent suborganellar localization analysis revealed a more prominent medial/trans-Golgi localization of NAA60. Our findings underscore the role of NAA60 and its interactors in maintaining Golgi structural integrity and highlight the effectiveness of BioID in generating critical protein topology data, invaluable for enhancing the prediction of protein topology within cellular compartments.}},
  articleno    = {{240225}},
  author       = {{Tanco, Sebastian and Jonckheere, Veronique and Tharkeshwar Raghunath, Arun Kumar and Bogaert, Annelies and Gevaert, Kris and Anneart, Wim and Van Damme, Petra}},
  issn         = {{2046-2441}},
  journal      = {{OPEN BIOLOGY}},
  keywords     = {{proximity-dependent biotin identification (BioID),Golgi fragmentation,NAA60,N-terminal acetylation,proxeome,transmembrane protein topology,ARF-LIKE GTPASE,ALPHA-ACETYLTRANSFERASE,MOLECULAR DETERMINANTS,ACETYLATION,IDENTIFICATION,TRAFFICKING,PLATFORM}},
  language     = {{eng}},
  number       = {{2}},
  pages        = {{16}},
  title        = {{Proximal partners of the organellar N-terminal acetyltransferase NAA60 : insights into Golgi structure and transmembrane protein topology}},
  url          = {{http://doi.org/10.1098/rsob.240225}},
  volume       = {{15}},
  year         = {{2025}},
}
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