title: In-gel N-acetylation for the quantification of the degree of protein in vivo N-terminal acetylation.
abstract: Maturation of protein N-termini occurs in all kingdoms of life, with major protein modifications being proteolytic processing (e.g., removal of initiator methionines) and N-terminal acetylation. The functional consequences of these modifications are only known for a few substrates, and techniques to study such modifications have begun to emerge only recently. We here report on a method enabling targeted, mass spectrometry based analysis of protein N-termini from polyacrylamide gel-separated proteins. In our method, stable isotope incorporation by in-gel N-acetylation of free primary amines permits calculating the extent of in vivo N-terminal acetylation, proven to reveal crucial information with reference to N-terminal protein biology.
authors: Department of Medical Protein Research, VIB, Ghent, Belgium.
authors: Petra
authors: P
authors: Van Damme
authors: Thomas
authors: T
authors: Arnesen
authors: Bart
authors: B
authors: Ruttens
authors: Kris
authors: K
authors: Gevaert
mesh_terms: D000107:Acetylation; D048868:Adaptor Proteins, Signal Transducing / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005782:Gels; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D063206:N-Terminal Acetyltransferases / Q000378:metabolism*; D009687:Nuclear Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D005782:Gels; D009687:Nuclear Proteins; C465618:PCNP protein, human; D011506:Proteins; D063206:N-Terminal Acetyltransferases