{"doi":"10.1186/1753-6561-3-s6-s5","title":"Application of reverse-phase HPLC to quantify oligopeptide acetylation eliminates interference from unspecific acetyl CoA hydrolysis","abstract":"<jats:title>Abstract</jats:title>\n          <jats:p>Protein acetylation is a common modification that plays a central role in several cellular processes. The most widely used methods to study these modifications are either based on the detection of radioactively acetylated oligopetide products or an enzyme-coupled reaction measuring conversion of the acetyl donor acetyl CoA to the product CoASH. Due to several disadvantages of these methods, we designed a new method to study oligopeptide acetylation. Based on reverse phase HPLC we detect both reaction products in a highly robust and reproducible way. The method reported here is also fully compatible with subsequent product analysis, e.g. by mass spectroscopy. The catalytic subunit, hNaa30p, of the human NatC protein N-acetyltransferase complex was used for N-terminal oligopeptide acetylation. We show that unacetylated and acetylated oligopeptides can be efficiently separated and quantified by the HPLC-based analysis. The method is highly reproducible and enables reliable quantification of both substrates and products. It is therefore well-suited to determine kinetic parameters of acetyltransferases.</jats:p>","journal":"BMC Proceedings","year":2009,"id":14585,"datarank":1.5061522438853878,"base_score":3.091042453358316,"endowment":3.091042453358316,"self_citation_contribution":0.4636563680037475,"citation_network_contribution":1.0424958758816403,"self_endowment_contribution":0.4636563680037475,"citer_contribution":1.0424958758816403,"corpus_percentile":null,"corpus_rank":null,"citation_count":21,"citer_count":15,"citers_with_citation_signal":14,"citers_with_endowment":14,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":114576,"name":"Kristine Hole","orcid":null,"position":1,"is_corresponding":false},{"id":114577,"name":"Mathias Ziegler","orcid":null,"position":2,"is_corresponding":false},{"id":114578,"name":"Johan R Lillehaug","orcid":null,"position":3,"is_corresponding":false},{"id":114575,"name":"Rune Evjenth","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"base_score":3.091042453358316,"endowment":3.091042453358316,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"19660098","pmcid":"PMC2722098","openalex_id":"https://openalex.org/W2150352169","authors":[],"funders":[{"funder_name":"NIGMS NIH HHS","grant_id":"R01 GM031575","title":null}],"total_grants":1,"fwci":1.0617,"citation_percentile":0.78388278,"influential_citations":0,"citation_trend":[{"year":2012,"count":4},{"year":2014,"count":2},{"year":2015,"count":3},{"year":2016,"count":1},{"year":2017,"count":1},{"year":2019,"count":1},{"year":2020,"count":1},{"year":2021,"count":1},{"year":2022,"count":1},{"year":2023,"count":1},{"year":2024,"count":1},{"year":2025,"count":1}],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://bmcproc.biomedcentral.com/counter/pdf/10.1186/1753-6561-3-S6-S5","host_type":"journal"},{"url":"https://bmcproc.biomedcentral.com/counter/pdf/10.1186/1753-6561-3-S6-S5","host_type":"GOLD"},{"url":"https://bmcproc.biomedcentral.com/counter/pdf/10.1186/1753-6561-3-S6-S5","host_type":"publisher"},{"url":"https://link.springer.com/content/pdf/10.1186/1753-6561-3-S6-S5.pdf","host_type":"publisher"},{"url":"https://doi.org/10.1186/1753-6561-3-s6-s5","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/19660098","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/2722098","host_type":"repository"},{"url":"http://www.biomedcentral.com/1753-6561/3_suppl_6/S5/abstract","host_type":"BioMedCentral"},{"url":"http://www.biomedcentral.com/content/pdf/1753-6561-3_suppl_6-S5.pdf","host_type":"BioMedCentral"},{"url":"http://www.biomedcentral.com/1753-6561/3_suppl_6/S5","host_type":"BioMedCentral"},{"url":"https://europepmc.org/articles/PMC2722098","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC2722098?pdf=render","host_type":"Europe_PMC"}],"fields_of_study":["Peptidase Inhibition and Analysis","Chemical Synthesis and Analysis","Neuropeptides and Animal Physiology","Chemistry","Medicine"],"mesh_terms":[],"keywords":["Acetylation","Acetyltransferase","Oligopeptide","Acetyltransferases","Acetyl-CoA","High-performance liquid chromatography","Chromatography","Chemistry","Hydrolysis","Biochemistry","Enzyme","Peptide"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"uniprot"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-06-01T12:30:30.835286Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}