title: Examining ubiquitinated peptide enrichment efficiency through an epitope labeled protein.
abstract: Ubiquitination is a dynamic process that is responsible for regulation of cellular responses to stimuli in a number of biological systems. Previous efforts to study this post-translational modification have focused on protein enrichment; however, recent research utilizes the presence of the di-glycine (Gly-Gly) remnants following trypsin digestion to immuno-enrich ubiquitinated peptides. Monoclonal antibodies developed to the cleaved ubiquitin modification epitope, (tert-butoxycarbonyl) glycylglycine (Boc-Gly-Gly-NHS)(1), are used to identify the Gly-Gly signature. Here, we have successfully generated the Boc-Gly-Gly-NHS modification and showed that when conjugated to a lysine containing protein, such as lysozyme, it can be applied as a standard protein to examine ubiquitinated peptide enrichment within a complex background.
authors: Department of Chemistry, North Carolina State University, Raleigh, NC, United States; W. M. Keck FTMS Laboratory for Human Health Research, North Carolina State University, Raleigh, NC, United States.
authors: J
authors: J
authors: Parker
authors: Department of Plant Pathology, North Carolina State University, Raleigh, NC, United States; Center for Integrated Fungal Research, North Carolina State University, Raleigh, NC, United States.
authors: Y
authors: Y
authors: Oh
authors: Department of Chemistry, North Carolina State University, Raleigh, NC, United States.
authors: Y
authors: Y
authors: Moazami
authors: Department of Chemistry, North Carolina State University, Raleigh, NC, United States.
authors: J G
authors: JG
authors: Pierce
authors: Department of Chemistry, North Carolina State University, Raleigh, NC, United States; W. M. Keck FTMS Laboratory for Human Health Research, North Carolina State University, Raleigh, NC, United States.
authors: P L
authors: PL
authors: Loziuk
authors: Department of Plant Pathology, North Carolina State University, Raleigh, NC, United States; Center for Integrated Fungal Research, North Carolina State University, Raleigh, NC, United States.
authors: R A
authors: RA
authors: Dean
authors: Department of Chemistry, North Carolina State University, Raleigh, NC, United States; W. M. Keck FTMS Laboratory for Human Health Research, North Carolina State University, Raleigh, NC, United States. Electronic address: dcmuddim@ncsu.edu.
authors: D C
authors: DC
authors: Muddiman
mesh_terms: D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry; D000939:Epitopes / Q000737:chemistry*; D009113:Muramidase / Q000737:chemistry*; D057149:Ubiquitinated Proteins / Q000737:chemistry*; D054875:Ubiquitination
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D000939:Epitopes; D057149:Ubiquitinated Proteins; D009113:Muramidase
keywords: Enrichment efficiency; Gly-Gly enrichment; Ubiquitination