Research Goals.
Our research seeks to delineate the structure->activity->function algorithm focusing on proteolytic enzymes as signaling elements. Our laboratory has very broad interests in principles of proteolysis in humans, and we take multi-pronged approaches to research on proteases and their inhibitors.
Regulated Cell Death
In one approach we apply basic biochemical knowledge to investigate newly emerging principles of proteolysis in human and mouse systems. This research is currently directed at dissecting the proteolytic components of the intracellular pathway that leads to regulated cell death, including apoptosis, pyroptosis, necroptosis, and necrosis. These programs contains a number of proteolytic steps that are essential for efficient execution of the death pathways. Thus the proteases of the pathway - the caspases, cathepsins, and hematopoietic serine proteases - are involved in the normal maintenance of correct cell number, and are therefore implicated in a number of pathologic and physiologic conditions. Using the techniques of protein chemistry, enzymology, crystallography, and genome editing, we analyze the basic mechanism utilized by proteases to promote cell death pathways, and the mechanisms and specificity of the natural inhibitors that control them. Key paper:
Salvesen GS, Hempel A, Coll NS.
Protease signaling in animal and plant-regulated cell death.
FEBS J. 2016 Jul;283(14):2577-98.
Cell Signaling
Modification of proteins by the small ubiquitin-like modifier SUMO is a dynamic and reversible process. The SUMO cycle begins when SUMO precursors are processed to remove short C-terminal extensions, thereby uncapping the C-terminal Gly-Gly motif that is essential for conjugation. SUMO ligases conjugate the protein, via its C-terminal carboxylate, to the side-chain lysine of target proteins to generate an isopeptide linkage. Eventually, SUMO is removed intact from its substrate SUMOylated proteins, and so the SUMOylation/deSUMOylation cycle regulates SUMOs function. A group of proteases known as SENPs are involved in both the activation of SUMO precursors (endopeptidase cleavage) and deconjugation of the targets (isopeptidase cleavage). Our laboratory seeks to define the mechanisms that mediate the regulation of SENP activity and access to their natural substrates. Key paper:
Fasci D, Anania VG, Lill JR, Salvesen GS.
SUMO deconjugation is required for arsenic-triggered ubiquitylation of PML.
Sci Signal. 2015 Jun 9;8(380)
Technology Development
The principle of proteolysis in vivo is to instigate irreversible changes to a set of protein substrates that alters their function, and generates the required biological event. The sum total of the proteases and their target substrates operating in a physiologic pathway therefore defines the global event. Consequently, the identity of the substrate cleavages defines the proteases acting on them. We are developing proteomics based methodologies, including selective protein labeling and mass spectrometry techniques, to identify the products of proteolysis in vivo. Key paper:
Timmer JC, Zhu W, Pop C, Regan T, Snipas SJ, Eroshkin AM, Riedl SJ, Salvesen GS.
Structural and kinetic determinants of protease substrates.
Nat Struct Mol Biol. 2009 Sep 20.
Chemical Biology
We collaborate extensively with the labs of Marcin Drag and Matt Bogyo to utilize peptide-based substrates and probes to define, visualize, and explore the role of cysteine and serine proteases in cell signaling. Key paper:
Kasperkiewicz P, Altman Y, D'Angelo M, Salvesen GS, Drag M.
Toolbox of Fluorescent Probes for Parallel Imaging Reveals Uneven Location of Serine Proteases in Neutrophils.
J Am Chem Soc. 2017 Jul 26;139(29):10115-10125.
Our research seeks to delineate the structure->activity->function algorithm focusing on proteolytic enzymes as signaling elements. Our laboratory has very broad interests in principles of proteolysis in humans, and we take multi-pronged approaches to research on proteases and their inhibitors.
Regulated Cell Death
In one approach we apply basic biochemical knowledge to investigate newly emerging principles of proteolysis in human and mouse systems. This research is currently directed at dissecting the proteolytic components of the intracellular pathway that leads to regulated cell death, including apoptosis, pyroptosis, necroptosis, and necrosis. These programs contains a number of proteolytic steps that are essential for efficient execution of the death pathways. Thus the proteases of the pathway - the caspases, cathepsins, and hematopoietic serine proteases - are involved in the normal maintenance of correct cell number, and are therefore implicated in a number of pathologic and physiologic conditions. Using the techniques of protein chemistry, enzymology, crystallography, and genome editing, we analyze the basic mechanism utilized by proteases to promote cell death pathways, and the mechanisms and specificity of the natural inhibitors that control them. Key paper:
Salvesen GS, Hempel A, Coll NS.
Protease signaling in animal and plant-regulated cell death.
FEBS J. 2016 Jul;283(14):2577-98.
Cell Signaling
Modification of proteins by the small ubiquitin-like modifier SUMO is a dynamic and reversible process. The SUMO cycle begins when SUMO precursors are processed to remove short C-terminal extensions, thereby uncapping the C-terminal Gly-Gly motif that is essential for conjugation. SUMO ligases conjugate the protein, via its C-terminal carboxylate, to the side-chain lysine of target proteins to generate an isopeptide linkage. Eventually, SUMO is removed intact from its substrate SUMOylated proteins, and so the SUMOylation/deSUMOylation cycle regulates SUMOs function. A group of proteases known as SENPs are involved in both the activation of SUMO precursors (endopeptidase cleavage) and deconjugation of the targets (isopeptidase cleavage). Our laboratory seeks to define the mechanisms that mediate the regulation of SENP activity and access to their natural substrates. Key paper:
Fasci D, Anania VG, Lill JR, Salvesen GS.
SUMO deconjugation is required for arsenic-triggered ubiquitylation of PML.
Sci Signal. 2015 Jun 9;8(380)
Technology Development
The principle of proteolysis in vivo is to instigate irreversible changes to a set of protein substrates that alters their function, and generates the required biological event. The sum total of the proteases and their target substrates operating in a physiologic pathway therefore defines the global event. Consequently, the identity of the substrate cleavages defines the proteases acting on them. We are developing proteomics based methodologies, including selective protein labeling and mass spectrometry techniques, to identify the products of proteolysis in vivo. Key paper:
Timmer JC, Zhu W, Pop C, Regan T, Snipas SJ, Eroshkin AM, Riedl SJ, Salvesen GS.
Structural and kinetic determinants of protease substrates.
Nat Struct Mol Biol. 2009 Sep 20.
Chemical Biology
We collaborate extensively with the labs of Marcin Drag and Matt Bogyo to utilize peptide-based substrates and probes to define, visualize, and explore the role of cysteine and serine proteases in cell signaling. Key paper:
Kasperkiewicz P, Altman Y, D'Angelo M, Salvesen GS, Drag M.
Toolbox of Fluorescent Probes for Parallel Imaging Reveals Uneven Location of Serine Proteases in Neutrophils.
J Am Chem Soc. 2017 Jul 26;139(29):10115-10125.
