Protein Termini Part A

by Thomas Arnesen (Editor)

Overview

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. - Provides the authority and expertise of leading contributors from an international board of authors - Includes the latest information on modifications and targeting of proteins via their N- and C-terminal ends - Presents a broad spectrum of methods within protein acetylation, myristoylation, arginylation and oxidation

Product Details

ISBN-13:	9780443414848
Publisher:	Elsevier Science & Technology Books
Publication date:	08/01/2025
Series:	Methods in Enzymology , #718
Sold by:	Barnes & Noble
Format:	eBook
Pages:	412
File size:	13 MB
Note:	This product may take a few minutes to download.

About the Author

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.

Table of Contents

1. In vitro acetyltransferase assays for NAT enzymesRonen Marmorstein2. A fluorescent CPM-based in vitro acetylation assay: a tool for assessing N-Terminal acetyltransferase activity and profiling compound activityTanja Bange3. Methods for Purification and Activity Analysis of N-Terminal AcetyltransferasesRong Huang4. N-terminal acetylation of plastid precursor proteinsSacha Baginsky5. Utilizing N-terminal acetylation specific antibodiesHenriette Aksnes6. Functional assessment of N-terminal acetyltransferase variantsNina McTiernan7. Affinity purification-mass spectrometry to identify nuclear protein interactions of N-terminal acetyltransferase NAA40Antonis Kirmizis8. A pan-N-formylmethionine-specific antibody as a tool to for analyzing Nα︎-terminal formylationCheol-Sang Hwang9. Chemical proteomic approaches to investigate N-myristoylationEdward W. Tate10. Analysis of the oxidation state of N-terminal methioninesSina Ghaemmaghami11. A biophysical approach to studying N-terminal cysteine oxidase substrate preferencesMark White12. Recombinant expression, purification, and characterization of human ATE1 arginyltransferaseYi Zhang13. In-bacteria arginylation assayYi Zhang14. Identification of arginylated N-termini with specific antibodiesJasper Eising15. Characterization of the autophagic N-degron pathway and monitoring its chemical modulation for therapeutic developmentYong Tae Kwon16. Developing a flow cytometric method to evaluate the stability of protein N-terminiAditya M. Kunjapur17. Degronopedia: A practical guide to identifying and targeting protein degronsWojciech Pokrzywa18. Generation and usage of ubiquitin-reference technique (URT) plasmids to validate N-degronsChang Hoon Ji19. Quantitative Insights into Protein Turnover and Ubiquitination with HiBiT and NanoBRETWojciech Pokrzywa20. Characterization of the autophagy E3 ligase/N-recognin KCMF1Chang Hoon Ji21. Identification of Ac/N-degron-recognition domain within the MARCHF6 E3 ubiquitin ligaseCheol-Sang Hwang22. TurboID technique for proximity labelling of interacting proteinsGreta Jarck23. Proteome analysis of nascent polypeptide chains using puromycinKoshi Imami24. HUNTER-DIA: An updated protocol for enrichment and mass spectrometry-based identification of protein N-terminiPitter F. Huesgen25. TERMINER - Bioinformatic detection and annotation of proteolytic protein termini in shotgun proteomics dataOliver Schilling26. Chemical proteomic approaches to investigate S-prenylationEdward W. Tate27. Quantitative analysis of C-terminal prenylated protein levels using tandem mass taggingMark Distefano28. Optimizing purification and FP-based binding assays for the E3 ligase FEM1CRong Huang

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Protein termini: key hubs for protein regulation