Detection and biochemical characterization of circulating proteasomes in dog plasma
A growing body of evidence convincingly indicates that proteasomes are not located exclusively within cells but also in different extracellular compartments. In humans, in fact, this large multimeric protease has been identified in many body fluids and secretions such as blood, urine, tears, sweat,...
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| Published in | Research in veterinary science Vol. 162; p. 104950 |
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| Main Authors | , , , , , |
| Format | Journal Article |
| Language | English |
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England
Elsevier Ltd
01.09.2023
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| Abstract | A growing body of evidence convincingly indicates that proteasomes are not located exclusively within cells but also in different extracellular compartments. In humans, in fact, this large multimeric protease has been identified in many body fluids and secretions such as blood, urine, tears, sweat, saliva, milk, and cerebrospinal and pericardial fluid. Intriguingly, the exact origins of these extracellular proteasomes as well as the specific biological functions they perform are largely unknown. As no data on this important subject is yet available in domestic animals, the present study was undertaken to investigate the presence of extracellular proteasomes in canine blood. As a result, for the first time, circulating proteasomes could be clearly detected in the plasma of a cohort of 20 healthy dogs. Furthermore, all three main proteasomal peptidase activities were measured and characterized using fluorogenic peptides and highly specific inhibitors. Finally, the effect of ATP and PA28 family activators on this circulating proteasome was investigated. Collectively, our data indicate that at least a part of the proteasome present in dog plasma consists of a particle that in vitro displays the enzymatic properties of the 20S proteasome.
•For the first time, circulating proteasomes were detected in dog plasma.•All three main proteasomal peptidase activities were measured and characterized.•Proteasomal activity detected in plasma is insensitive to ATP.•Proteasomal activity detected in dog plasma is highly enhanced by PA28αβ and γ. |
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| AbstractList | A growing body of evidence convincingly indicates that proteasomes are not located exclusively within cells but also in different extracellular compartments. In humans, in fact, this large multimeric protease has been identified in many body fluids and secretions such as blood, urine, tears, sweat, saliva, milk, and cerebrospinal and pericardial fluid. Intriguingly, the exact origins of these extracellular proteasomes as well as the specific biological functions they perform are largely unknown. As no data on this important subject is yet available in domestic animals, the present study was undertaken to investigate the presence of extracellular proteasomes in canine blood. As a result, for the first time, circulating proteasomes could be clearly detected in the plasma of a cohort of 20 healthy dogs. Furthermore, all three main proteasomal peptidase activities were measured and characterized using fluorogenic peptides and highly specific inhibitors. Finally, the effect of ATP and PA28 family activators on this circulating proteasome was investigated. Collectively, our data indicate that at least a part of the proteasome present in dog plasma consists of a particle that in vitro displays the enzymatic properties of the 20S proteasome. A growing body of evidence convincingly indicates that proteasomes are not located exclusively within cells but also in different extracellular compartments. In humans, in fact, this large multimeric protease has been identified in many body fluids and secretions such as blood, urine, tears, sweat, saliva, milk, and cerebrospinal and pericardial fluid. Intriguingly, the exact origins of these extracellular proteasomes as well as the specific biological functions they perform are largely unknown. As no data on this important subject is yet available in domestic animals, the present study was undertaken to investigate the presence of extracellular proteasomes in canine blood. As a result, for the first time, circulating proteasomes could be clearly detected in the plasma of a cohort of 20 healthy dogs. Furthermore, all three main proteasomal peptidase activities were measured and characterized using fluorogenic peptides and highly specific inhibitors. Finally, the effect of ATP and PA28 family activators on this circulating proteasome was investigated. Collectively, our data indicate that at least a part of the proteasome present in dog plasma consists of a particle that in vitro displays the enzymatic properties of the 20S proteasome.A growing body of evidence convincingly indicates that proteasomes are not located exclusively within cells but also in different extracellular compartments. In humans, in fact, this large multimeric protease has been identified in many body fluids and secretions such as blood, urine, tears, sweat, saliva, milk, and cerebrospinal and pericardial fluid. Intriguingly, the exact origins of these extracellular proteasomes as well as the specific biological functions they perform are largely unknown. As no data on this important subject is yet available in domestic animals, the present study was undertaken to investigate the presence of extracellular proteasomes in canine blood. As a result, for the first time, circulating proteasomes could be clearly detected in the plasma of a cohort of 20 healthy dogs. Furthermore, all three main proteasomal peptidase activities were measured and characterized using fluorogenic peptides and highly specific inhibitors. Finally, the effect of ATP and PA28 family activators on this circulating proteasome was investigated. Collectively, our data indicate that at least a part of the proteasome present in dog plasma consists of a particle that in vitro displays the enzymatic properties of the 20S proteasome. A growing body of evidence convincingly indicates that proteasomes are not located exclusively within cells but also in different extracellular compartments. In humans, in fact, this large multimeric protease has been identified in many body fluids and secretions such as blood, urine, tears, sweat, saliva, milk, and cerebrospinal and pericardial fluid. Intriguingly, the exact origins of these extracellular proteasomes as well as the specific biological functions they perform are largely unknown. As no data on this important subject is yet available in domestic animals, the present study was undertaken to investigate the presence of extracellular proteasomes in canine blood. As a result, for the first time, circulating proteasomes could be clearly detected in the plasma of a cohort of 20 healthy dogs. Furthermore, all three main proteasomal peptidase activities were measured and characterized using fluorogenic peptides and highly specific inhibitors. Finally, the effect of ATP and PA28 family activators on this circulating proteasome was investigated. Collectively, our data indicate that at least a part of the proteasome present in dog plasma consists of a particle that in vitro displays the enzymatic properties of the 20S proteasome. •For the first time, circulating proteasomes were detected in dog plasma.•All three main proteasomal peptidase activities were measured and characterized.•Proteasomal activity detected in plasma is insensitive to ATP.•Proteasomal activity detected in dog plasma is highly enhanced by PA28αβ and γ. |
| ArticleNumber | 104950 |
| Author | Mengozzi, G. Borrelli, A. Cascio, P. Borella, F. Degiovanni, A. Cerruti, F. |
| Author_xml | – sequence: 1 givenname: F. surname: Cerruti fullname: Cerruti, F. organization: Department of Veterinary Sciences, University of Turin, Largo P. Braccini, 2, 10095, Grugliasco, Turin, Italy – sequence: 2 givenname: A. surname: Borrelli fullname: Borrelli, A. organization: Department of Veterinary Sciences, University of Turin, Largo P. Braccini, 2, 10095, Grugliasco, Turin, Italy – sequence: 3 givenname: A. surname: Degiovanni fullname: Degiovanni, A. organization: Department of Veterinary Sciences, University of Turin, Largo P. Braccini, 2, 10095, Grugliasco, Turin, Italy – sequence: 4 givenname: G. surname: Mengozzi fullname: Mengozzi, G. organization: Department of Public Health and Pediatric Sciences, University of Turin, C.so Bramante, 88/90, 10100 Turin, Italy – sequence: 5 givenname: F. surname: Borella fullname: Borella, F. organization: Department of Veterinary Sciences, University of Turin, Largo P. Braccini, 2, 10095, Grugliasco, Turin, Italy – sequence: 6 givenname: P. surname: Cascio fullname: Cascio, P. email: paolo.cascio@unito.it organization: Department of Veterinary Sciences, University of Turin, Largo P. Braccini, 2, 10095, Grugliasco, Turin, Italy |
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| Keywords | Circulating 20S Proteasomal activities Canine plasma Proteasome activators Extracellular proteasome |
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| Title | Detection and biochemical characterization of circulating proteasomes in dog plasma |
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