Protein Termini Part A

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Thomas Arnesen (Researcher, Professor, The Department of Biomedicine, Department of Biological Sciences (BIO), Arnesen Lab, University of Bergen, Bergen, Norway)

Protein Termini Part A

Thomas Arnesen (Researcher, Professor, The Department of Biomedicine, Department of Biological Sciences (BIO), Arnesen Lab, University of Bergen, Bergen, Norway)

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English

Academic Press Inc
01 October 2025

Biophysics; Biochemistry; Molecular biology

Series: Methods in Enzymology

Protein termini represent a major route to protein regulation. From the moment the very first amino acid of a polypeptide chain exits the ribosome there is potential for steering from the cellular environment. This volume of Methods in Enzymology Modifications and Targeting of Protein Termini focuses on Protein N-termini and C-termini and their modifications which include acetylation, arginylation, myristoylation and oxidation. Also, the impact of terminal modifications is covered, in particular the impact on protein turnover and the ubiquitin E3 ligases which specifically recognize protein N-termini (N-degrons) and C-termini (C-degrons). In addition to the detailed methods and laboratory protocols, the chapters include informative overviews and reviews of the different subfields.

Volume editor: Thomas Arnesen (Researcher Professor The Department of Biomedicine Department of Biological Sciences (BIO) Arnesen Lab University of Bergen Bergen Norway)
Imprint: Academic Press Inc
Country of Publication: United States
Dimensions: Height: 229mm, Width: 152mm,
Weight: 450g
ISBN: 9780443414831
ISBN 10: 0443414831
Series: Methods in Enzymology
Pages: 412
Publication Date: 01 October 2025
Audience: Professional and scholarly , Undergraduate
Format: Hardback
Publisher's Status: Active

1. In vitro acetyltransferase assays for NAT enzymes Ronen Marmorstein 2. A fluorescent CPM-based in vitro acetylation assay: a tool for assessing N-Terminal acetyltransferase activity and profiling compound activity Tanja Bange 3. Methods for Purification and Activity Analysis of N-Terminal Acetyltransferases Rong Huang 4. N-terminal acetylation of plastid precursor proteins Sacha Baginsky 5. Utilizing N-terminal acetylation specific antibodies Henriette Aksnes 6. Functional assessment of N-terminal acetyltransferase variants Nina McTiernan 7. Affinity purification-mass spectrometry to identify nuclear protein interactions of N-terminal acetyltransferase NAA40 Antonis Kirmizis 8. A pan-N-formylmethionine-specific antibody as a tool to for analyzing Nα︎-terminal formylation Cheol-Sang Hwang 9. Chemical proteomic approaches to investigate N-myristoylation Edward W. Tate 10. Analysis of the oxidation state of N-terminal methionines Sina Ghaemmaghami 11. A biophysical approach to studying N-terminal cysteine oxidase substrate preferences Mark White 12. Recombinant expression, purification, and characterization of human ATE1 arginyltransferase Yi Zhang 13. In-bacteria arginylation assay Yi Zhang 14. Identification of arginylated N-termini with specific antibodies Jasper Eising 15. Characterization of the autophagic N-degron pathway and monitoring its chemical modulation for therapeutic development Yong Tae Kwon 16. Developing a flow cytometric method to evaluate the stability of protein N-termini Aditya M. Kunjapur 17. Degronopedia: A practical guide to identifying and targeting protein degrons Wojciech Pokrzywa 18. Generation and usage of ubiquitin-reference technique (URT) plasmids to validate N-degrons Chang Hoon Ji 19. Quantitative Insights into Protein Turnover and Ubiquitination with HiBiT and NanoBRET Wojciech Pokrzywa 20. Characterization of the autophagy E3 ligase/N-recognin KCMF1 Chang Hoon Ji 21. Identification of Ac/N-degron-recognition domain within the MARCHF6 E3 ubiquitin ligase Cheol-Sang Hwang 22. TurboID technique for proximity labelling of interacting proteins Greta Jarck 23. Proteome analysis of nascent polypeptide chains using puromycin Koshi Imami 24. HUNTER-DIA: An updated protocol for enrichment and mass spectrometry-based identification of protein N-termini Pitter F. Huesgen 25. TERMINER - Bioinformatic detection and annotation of proteolytic protein termini in shotgun proteomics data Oliver Schilling 26. Chemical proteomic approaches to investigate S-prenylation Edward W. Tate 27. Quantitative analysis of C-terminal prenylated protein levels using tandem mass tagging Mark Distefano 28. Optimizing purification and FP-based binding assays for the E3 ligase FEM1C Rong Huang

Professor Thomas Arnesen received his Ph.D. in molecular biology from the University of Bergen, Norway in 2006. After postdoctoral work at Haukeland University Hospital and University of Rochester Medical Center, he established his own lab at the University of Bergen in 2010. His main interest has been protein N-terminal acetylation and the responsible enzymes, the N-terminal acetyltransferases (NATs). Using Saccharomyces cerevisiae and human cell models combined with in vitro approaches his lab and collaborators have i) identified and defined the presumed complete cytosolic human NAT-machinery including NATs acting post-translationally, ii) quantitatively analysed the N-terminal acetylomes of yeast and human cells, iii) developed novel assays for NAT-profiling, iv) gained mechanistic insights of the molecular and cellular effects of N-terminal acetylation, v) contributed to the understanding of the physiological and clinical importance of NATs by revealing the links between NatA and cancer cell survival and drug sensitisation, and lately by defining genetic disorders caused by pathogenic NAT variants. The Arnesen lab also contributed to solving the first NAT-structures and developing the first potent NAT-inhibitors. With Fred Sherman and Bogdan Polevoda, Arnesen introduced the NAA (N-alpha acetyltransferase) nomenclature of the N-terminal acetyltransferase genes and proteins, and he acts as the specialist advisor for the HUGO Gene Nomenclature Committee for these genes. Arnesen is one of the founders and council members of the International Society of Protein Termini (ISPT). He has organized several symposia on N-terminal acetylation, and in 2022 he was the head organizer of the EMBO Workshop ‘Protein Termini – From mechanism to biological impact’ in Bergen, Norway. Arnesen has co-authored more than 100 peer-reviewed publications. Today he is head of the Translational Cell Signaling and Metabolism group at the Dept. of Biomedicine, UiB, supported by the Research Council of Norway and ERC. Here his team continues the basic and translational research to understand the impact of protein N-terminal modifications.