Title

Structural analysis of a highly acetylated protein using a curved-field reflectron mass spectrometer

UMMS Affiliation

Department of Biochemistry and Molecular Pharmacology

Date

6-1-2005

Document Type

Article

Medical Subject Headings

Acetylation; Amino Acid Sequence; Binding Sites; Cell Cycle Proteins; Histone Acetyltransferases; Lysine; Molecular Sequence Data; Peptides; Recombinant Proteins; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Transcription Factors; p300-CBP Transcription Factors

Disciplines

Biochemistry | Enzymes and Coenzymes | Medicinal-Pharmaceutical Chemistry | Therapeutics

Abstract

Matrix-assisted laser desorption/ionization mass spectrometry and tandem mass spectrometry (MS/MS) were used to determine the multiple acetylation sites in the histone acetyltransferase (HAT): p300-HAT. Partial cleavage of the peptides containing acetylated lysine residues by trypsin provided a set of nested sequences that enabled us to determine that multiple acetylation occurs on the same molecule. At the same time, cleavages resulting in a terminal unacetylated lysine suggested that not all of these sites are fully modified. Using MS and MS/MS, we were able to characterize both the unmodified and acetylated tryptic peptides covering more than 82% of the protein.

Rights and Permissions

Citation: Proteomics. 2005 Jun;5(9):2288-96. Link to article on publisher's site

Comments

At the time of publication, Paul Thompson was not yet affiliated with UMass Medical School.

Related Resources

Link to Article in PubMed

Keywords

Acetylation, Amino acid sequencing, Matrix-assisted laser desorption/ionization-time of flight, Mass spectrometry