Title

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

UMMS Affiliation

Department of Biochemistry and Molecular Pharmacology

Publication Date

6-1-2005

Document Type

Article

Subjects

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.

Keywords

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

DOI of Published Version

10.1002/pmic.200401167

Source

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

Journal/Book/Conference Title

Proteomics

Comments

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

Related Resources

Link to Article in PubMed

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