Human cathepsins K, L, and S: Related proteases, but unique fibrinolytic activity

Douglas, Simone A.; Lamothe, Sarah E.; Singleton, Tatiyanna S.; Averett, Rodney D.; Platt, Manu O.

doi:10.1016/j.bbagen.2018.06.015

Cited by 14 publications

(10 citation statements)

References 46 publications

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“…In addition, our validation experiments corroborated the overexpression of CATS in an independent cohort of donors at day 21. This supports the theory by Douglas et al 31 that suggests cathepsins are involved in the degradation of a fibrin network with endothelial cells and fibroblast encapsulated, which collapsed in an unpredictable an uncontrolled manner in days to weeks 32 . In addition to their fibrinolytic properties, CATS also participates in healing processes.…”

Section: Discussionsupporting

confidence: 90%

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Deciphering the secretome of leukocyte-platelet rich fibrin: towards a better understanding of its wound healing properties

Hermida-Nogueira

Barrachina

Morán

et al. 2020

Sci Rep

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Leukocyte-platelet rich fibrin (L-PRF) is extensively used in the dentistry field and other clinical scenarios due to its regeneration properties. the goal of the present study was to depict the L-pRf secretome and how it changes over time. We obtained L-PRF membranes and cultured them in DMEM. The secretome was collected at days 3, 7 and 21. The secretome at day 3 was analysed by LC-MS/ MS and differences over time were analysed by Sequential Window Acquisition of all Theoretical Mass Spectra (SWATH). Overall, 705 proteins were identified in the secretome of L-PRF membranes after 3 days of culture, including growth factors (EGF, PDGFA) and proteins related to platelet and neutrophil degranulation. A total of 202 differentially secreted proteins were quantified by SWATH when comparing secretomes at days 3, 7 and 21. Most of them were enriched at day 3 such as MMP9, TSP1 and CO3. On the contrary, fibrinogen and CATS were found down-regulated at day 3. Growth factor and western blotting analysis corroborated the proteomic results. This is the most detailed proteome analysis of the L-PRF secretome to date. Proteins and growth factors identified, and their kinetics, provide novel information to further understand the wound healing properties of L-pRf. In the last two decades, the use of platelet-rich concentrates (PRC) has become very popular in different fields, especially in dentistry, due to their regeneration properties. Different scientists and companies have developed methods to obtained PRC with presence or absence of leukocytes but all of them with addition of anticoagulants to the tubes. In 2001, Choukroun et al. developed the second generation of PRC, Leukocyte platelet rich fibrin (L-PRF) 1 , which is obtained by blood centrifugation without anticoagulant in the tubes. Nowadays, L-PRF is the most commonly used PRC since it is easy to obtain and can be applied as a gel or compressed into a membrane depending on the application site. It has been mostly used in a dentistry field, especially for the treatment of osteonecrosis 2,3 , implant placement in bone-deficient sites 4 and sinus floor elevation 5. Additionally, L-PRF has started to be used in other clinical scenarios, for example for the treatment of chronic wounds, leg ulcers or diabetic foot 6 or for the treatment of ocular lesions 7. Wound healing is a complex process, which encompass different phases: haemostasis, inflammation, proliferation and remodelling. In each phase, different cell types take part in order to regenerate the tissue. Two types of blood cells, platelets and leukocytes, are among the cells that participate first. Growth factors, cytokines and other proteins released by these blood cells have chemoattractant properties that contribute to wound repair 8-10 .

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Section: Discussionsupporting

confidence: 90%

“…A recent study has demonstrated its role to hydrolyze the α and β chains of fibrin. Indeed, it was suggested the possibility of multiple sites of cleavage along fibrin because CATS can even cleave fragments produced after plasmin-mediated fibrinolysis 31 .…”

Section: Discussionmentioning

confidence: 99%

Deciphering the secretome of leukocyte-platelet rich fibrin: towards a better understanding of its wound healing properties

Hermida-Nogueira

Barrachina

Morán

et al. 2020

Sci Rep

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“…Cathepsins can be associated or bound to the ECM by a number of other noncovalent interactions that retain its associations with the protein even in the denaturing conditions of electrophoresis. We have demonstrated this with fibrin(ogen) binding of cathepsin L 59 , and there has even been shown that cathepsin-cystatin binding is sustained under SDS-PAGE as well 62 .…”

Section: Discussionmentioning

confidence: 71%

“…Studies have been published on the modifications of cathepsin activity by glycosaminoglycans, particularly in acidic conditions 32,57,58 . We and others have published that cathepsins are active by zymography while still associated with substrate 7,21,59–61 . Cathepsins can be associated or bound to the ECM by a number of other noncovalent interactions that retain its associations with the protein even in the denaturing conditions of electrophoresis.…”

Section: Discussionmentioning

confidence: 97%

Sequential, but not Concurrent, Incubation of Cathepsin K and L with Type I Collagen Results in Extended Proteolysis

Parks

Nahata

Edouard

et al. 2019

Sci Rep

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Degradation of extracellular matrix (ECM) during tendinopathy is, in part, mediated by the collagenolytic cathepsin K (catK) and cathepsin L (catL), with a temporal component to their activity. The objective of this study was to determine how catK and catL act in concert or in conflict to degrade collagen and tendon ECM during tissue degeneration. To do so, type I collagen gels or ECM extracted from apolipoprotein E deficient mouse Achilles tendons were incubated with catK and catL either concurrently or sequentially, incubating catK first, then catL after a delayed time period. Sequential incubation of catK then catL caused greater degradation of substrates over concurrent incubation, and of either cathepsin alone. Zymography showed there were reduced amounts of active enzymes when co-incubated, indicating that cannibalism, or protease-on-protease degradation between catK and catL was occurring, but incubation with ECM could distract from these interactions. CatK alone was sufficient to quickly degrade tendon ECM, but catL was not, requiring the presence of catK for degradation. Together, these data identify cooperative and conflicting actions of cathepsin mediated collagen matrix degradation by considering interactive effects of multiple proteases during tissue degeneration.

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“…Caspases are another system of proteases that cleave procaspases to the active caspase forms as a part of the apoptotic cascade, which then go and proteolyze their cellular targets (56). Cysteine cathepsins contribute to each of these functional responses (57,58), so there also may be interproteolytic family interactions in a more complex proteolytic network. For cysteine cathepsins, mature cathepsins cleave the propeptide from another procathepsin, activating it to the mature form.…”

Section: Discussionmentioning

confidence: 99%

Reassessing enzyme kinetics: Considering protease-as-substrate interactions in proteolytic networks

Ferrall‐Fairbanks

Kieslich

Platt

2020

Proc. Natl. Acad. Sci. U.S.A.

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Enzymes are catalysts in biochemical reactions that, by definition, increase rates of reactions without being altered or destroyed. However, when that enzyme is a protease, a subclass of enzymes that hydrolyze other proteins, and that protease is in a multiprotease system, protease-as-substrate dynamics must be included, challenging assumptions of enzyme inertness, shifting kinetic predictions of that system. Protease-on-protease inactivating hydrolysis can alter predicted protease concentrations used to determine pharmaceutical dosing strategies. Cysteine cathepsins are proteases capable of cathepsin cannibalism, where one cathepsin hydrolyzes another with substrate present, and misunderstanding of these dynamics may cause miscalculations of multiple proteases working in one proteolytic network of interactions occurring in a defined compartment. Once rates for individual protease-on-protease binding and catalysis are determined, proteolytic network dynamics can be explored using computational models of cooperative/competitive degradation by multiple proteases in one system, while simultaneously incorporating substrate cleavage. During parameter optimization, it was revealed that additional distraction reactions, where inactivated proteases become competitive inhibitors to remaining, active proteases, occurred, introducing another network reaction node. Taken together, improved predictions of substrate degradation in a multiple protease network were achieved after including reaction terms of autodigestion, inactivation, cannibalism, and distraction, altering kinetic considerations from other enzymatic systems, since enzyme can be lost to proteolytic degradation. We compiled and encoded these dynamics into an online platform (https://plattlab.shinyapps.io/catKLS/) for individual users to test hypotheses of specific perturbations to multiple cathepsins, substrates, and inhibitors, and predict shifts in proteolytic network reactions and system dynamics.

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Human cathepsins K, L, and S: Related proteases, but unique fibrinolytic activity

Abstract: Demonstration of cathepsins K, L, and S fibrinolytic activity leads to further investigation of contributory roles in disrupting vascular hemostasis, or breakdown of fibrin-based engineered vascular constructs where non-plasmin mediated fibrinolysis must be considered.

Cited by 14 publications

References 46 publications

Deciphering the secretome of leukocyte-platelet rich fibrin: towards a better understanding of its wound healing properties

Deciphering the secretome of leukocyte-platelet rich fibrin: towards a better understanding of its wound healing properties

Sequential, but not Concurrent, Incubation of Cathepsin K and L with Type I Collagen Results in Extended Proteolysis

Reassessing enzyme kinetics: Considering protease-as-substrate interactions in proteolytic networks

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