47 research outputs found
Identification of β-Secretase (BACE1) Substrates Using Quantitative Proteomics
β-site APP cleaving enzyme 1 (BACE1) is a transmembrane aspartyl protease with a lumenal active site that sheds the ectodomains of membrane proteins through juxtamembrane proteolysis. BACE1 has been studied principally for its role in Alzheimer's disease as the β-secretase responsible for generating the amyloid-β protein. Emerging evidence from mouse models has identified the importance of BACE1 in myelination and cognitive performance. However, the substrates that BACE1 processes to regulate these functions are unknown, and to date only a few β-secretase substrates have been identified through candidate-based studies. Using an unbiased approach to substrate identification, we performed quantitative proteomic analysis of two human epithelial cell lines stably expressing BACE1 and identified 68 putative β-secretase substrates, a number of which we validated in a cell culture system. The vast majority were of type I transmembrane topology, although one was type II and three were GPI-linked proteins. Intriguingly, a preponderance of these proteins are involved in contact-dependent intercellular communication or serve as receptors and have recognized roles in the nervous system and other organs. No consistent sequence motif predicting BACE1 cleavage was identified in substrates versus non-substrates. These findings expand our understanding of the proteins and cellular processes that BACE1 may regulate, and suggest possible mechanisms of toxicity arising from chronic BACE1 inhibition
Phosphoproteomic characterization of DNA damage response in melanoma cells following MEK/PI3K dual inhibition
Analysis of prerequisites violations financial stability
Світова економічна криза 2007–2008 років і потрясіння, що охо-
пили одночасно секторальні ринки кредитування, страхування, нерухомості та цінних паперів, продемонстрували, що системні ризики
підтримки фінансової стабільності не були належним чином оцінені
регуляторами
Complementary Proteomic Tools for the Dissection of Apoptotic Proteolysis Events
Proteolysis is a key regulatory event that controls intracellular
and extracellular signaling through irreversible changes in a protein’s
structure that greatly alters its function. Here we describe a platform
for profiling caspase substrates which encompasses two highly complementary
proteomic techniquesthe first is a differential gel based
approach termed Global Analyzer of SILAC-derived Substrates of Proteolysis
(GASSP) and the second involves affinity enrichment of peptides containing
a C-terminal aspartic acid residue. In combination, these techniques
have enabled the profiling of a large cellular pool of apoptotic-mediated
proteolytic events across a wide dynamic range. By applying this integrated
proteomic work flow to analyze proteolytic events resulting from the
induction of intrinsic apoptosis in Jurkat cells via etoposide treatment,
3346 proteins were quantified, of which 360 proteins were identified
as etoposide-induced proteolytic substrates, including 160 previously
assigned caspase substrates. In addition to global profiling, a targeted
approach using BAX HCT116 isogenic cell lines was utilized to dissect
pre- and post-mitochondrial extrinsic apoptotic cleavage events. By
employing apoptotic activation with a pro-apoptotic receptor agonist
(PARA), a limited set of apoptotic substrates including known caspase
substrates such as BH3 interacting-domain death agonist (BID) and
Poly (ADP-ribose) polymerase (PARP)-1, and novel substrates such as
Basic Transcription Factor 3, TRK-fused gene protein (TFG), and p62/Sequestosome
were also identified
Use of Differential Isotopic Labeling and Mass Spectrometry To Analyze Capacitation-Associated Changes in the Phosphorylation Status of Mouse Sperm Proteins
The Impact of Peptide Abundance and Dynamic Range on Stable-Isotope-Based Quantitative Proteomic Analyses
Complementary Proteomic Tools for the Dissection of Apoptotic Proteolysis Events
Proteolysis is a key regulatory event that controls intracellular
and extracellular signaling through irreversible changes in a protein’s
structure that greatly alters its function. Here we describe a platform
for profiling caspase substrates which encompasses two highly complementary
proteomic techniquesthe first is a differential gel based
approach termed Global Analyzer of SILAC-derived Substrates of Proteolysis
(GASSP) and the second involves affinity enrichment of peptides containing
a C-terminal aspartic acid residue. In combination, these techniques
have enabled the profiling of a large cellular pool of apoptotic-mediated
proteolytic events across a wide dynamic range. By applying this integrated
proteomic work flow to analyze proteolytic events resulting from the
induction of intrinsic apoptosis in Jurkat cells via etoposide treatment,
3346 proteins were quantified, of which 360 proteins were identified
as etoposide-induced proteolytic substrates, including 160 previously
assigned caspase substrates. In addition to global profiling, a targeted
approach using BAX HCT116 isogenic cell lines was utilized to dissect
pre- and post-mitochondrial extrinsic apoptotic cleavage events. By
employing apoptotic activation with a pro-apoptotic receptor agonist
(PARA), a limited set of apoptotic substrates including known caspase
substrates such as BH3 interacting-domain death agonist (BID) and
Poly (ADP-ribose) polymerase (PARP)-1, and novel substrates such as
Basic Transcription Factor 3, TRK-fused gene protein (TFG), and p62/Sequestosome
were also identified
Complementary Proteomic Tools for the Dissection of Apoptotic Proteolysis Events
Proteolysis is a key regulatory event that controls intracellular
and extracellular signaling through irreversible changes in a protein’s
structure that greatly alters its function. Here we describe a platform
for profiling caspase substrates which encompasses two highly complementary
proteomic techniquesthe first is a differential gel based
approach termed Global Analyzer of SILAC-derived Substrates of Proteolysis
(GASSP) and the second involves affinity enrichment of peptides containing
a C-terminal aspartic acid residue. In combination, these techniques
have enabled the profiling of a large cellular pool of apoptotic-mediated
proteolytic events across a wide dynamic range. By applying this integrated
proteomic work flow to analyze proteolytic events resulting from the
induction of intrinsic apoptosis in Jurkat cells via etoposide treatment,
3346 proteins were quantified, of which 360 proteins were identified
as etoposide-induced proteolytic substrates, including 160 previously
assigned caspase substrates. In addition to global profiling, a targeted
approach using BAX HCT116 isogenic cell lines was utilized to dissect
pre- and post-mitochondrial extrinsic apoptotic cleavage events. By
employing apoptotic activation with a pro-apoptotic receptor agonist
(PARA), a limited set of apoptotic substrates including known caspase
substrates such as BH3 interacting-domain death agonist (BID) and
Poly (ADP-ribose) polymerase (PARP)-1, and novel substrates such as
Basic Transcription Factor 3, TRK-fused gene protein (TFG), and p62/Sequestosome
were also identified
Complementary Proteomic Tools for the Dissection of Apoptotic Proteolysis Events
Proteolysis is a key regulatory event that controls intracellular
and extracellular signaling through irreversible changes in a protein’s
structure that greatly alters its function. Here we describe a platform
for profiling caspase substrates which encompasses two highly complementary
proteomic techniquesthe first is a differential gel based
approach termed Global Analyzer of SILAC-derived Substrates of Proteolysis
(GASSP) and the second involves affinity enrichment of peptides containing
a C-terminal aspartic acid residue. In combination, these techniques
have enabled the profiling of a large cellular pool of apoptotic-mediated
proteolytic events across a wide dynamic range. By applying this integrated
proteomic work flow to analyze proteolytic events resulting from the
induction of intrinsic apoptosis in Jurkat cells via etoposide treatment,
3346 proteins were quantified, of which 360 proteins were identified
as etoposide-induced proteolytic substrates, including 160 previously
assigned caspase substrates. In addition to global profiling, a targeted
approach using BAX HCT116 isogenic cell lines was utilized to dissect
pre- and post-mitochondrial extrinsic apoptotic cleavage events. By
employing apoptotic activation with a pro-apoptotic receptor agonist
(PARA), a limited set of apoptotic substrates including known caspase
substrates such as BH3 interacting-domain death agonist (BID) and
Poly (ADP-ribose) polymerase (PARP)-1, and novel substrates such as
Basic Transcription Factor 3, TRK-fused gene protein (TFG), and p62/Sequestosome
were also identified