Sensitive substrates for human leukocyte and porcine pancreatic elastase: A study of the merits of various chromophoric and fluorogenic leaving groups in assays for serine proteases

Castillo, Mario J.; Nakajima, Kiichiro; Zimmerman, M.D.; Powers, James C.

doi:10.1016/0003-2697(79)90043-5

Cited by 245 publications

(113 citation statements)

References 28 publications

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“…Thus, our study has expanded the list of pathogens (mycoplasma in addition to Gram-positive and Gram-negative bacteria) that may utilize IL-23/IL-17 axis to initiate a robust host defense against the invading microorganisms Here, a trend toward an increased lung Mp load, lower neutrophil content in lung tissue as well as in BAL reflected by MPO activity levels was observed in mice with IL-23 signaling being blocked. As a significant correlation between tissue MPO levels and neutrophil accumulation is well documented [20,[31][32][33][34][35], the decreased MPO activity levels due to IL-23 blocking suggest less neutrophil accumulation in the lung with reduced IL-17/IL-17F. In addition, although an abrogation of Mp-induced IL-17 /IL-17F and significantly decreased neutrophil levels were seen in the lung after IL-23 blocking, there was only a good, but not significant trend toward an increase of lung Mp load in vivo.…”

Section: Discussionmentioning

confidence: 99%

IL-23-dependent IL-17 production is essential in neutrophil recruitment and activity in mouse lung defense against respiratory Mycoplasma pneumoniae infection

Martin

Rino

et al. 2007

Microbes and Infection

250

185

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IL-23 induces IL-17 production in activated CD4+ T cells and participates in host defense against many encapsulated bacteria. However, whether IL-23/IL-17 axis contributes to a Mycoplasma pneumoniae (Mp)-induced lung inflammation (e.g., neutrophils) has not been addressed. Using an acute respiratory Mp infection murine model, we found significantly up-regulated lung IL-23p19 mRNA in the early phase of infection (4 h), and alveolar macrophages were an important cell source of Mp-induced IL-23. We further showed that Mp significantly increased IL-17 protein levels in bronchoalveolar lavage (BAL). Lung gene expression of IL-17, IL-17C and IL-17F was also markedly up-regulated by Mp in vivo. IL-17 and IL-17F were found to be derived mainly from lung CD4+ T cells, and were increased upon IL-23 stimulation in vitro. In vivo blocking of IL-23p19 alone or in combination with IL-23/IL-12p40 resulted in a significant reduction of Mp-induced IL-17 protein and IL-17/IL-17F mRNA expression, which was accompanied by a trend toward reduced lung neutrophil recruitment, BAL neutrophil activity, and Mp clearance. However, IL-23 neutralization had no effect on Mp-induced lung IL-17C mRNA expression. These results demonstrate that IL-17/IL-17F production is IL-23-dependent in an acute Mp infection, and contributes to neutrophil recruitment and activity in lung defense against the infection.

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Section: Discussionmentioning

confidence: 99%

IL-23-dependent IL-17 production is essential in neutrophil recruitment and activity in mouse lung defense against respiratory Mycoplasma pneumoniae infection

Martin

Rino

et al. 2007

Microbes and Infection

250

185

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“…Both paranitroaniline (pNA) and coumadinamide (CMA) have been used to assess elastase substrate activity through colorimetric and fluorescence detection [63]. Although the pNA substrate is highly specific for elastase the AMC substrate is much more sensitive.…”

Section: Cotton Cellulose Nanocrystals As Hne Sensorsmentioning

confidence: 99%

Nanocellulose-Based Biosensors: Design, Preparation, and Activity of Peptide-Linked Cotton Cellulose Nanocrystals Having Fluorimetric and Colorimetric Elastase Detection Sensitivity

Edwards¹,

Prevost²,

French³

et al. 2013

ENG

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Nanocrystalline cellulose is an amphiphilic, high surface area material that can be easily functionalized and is biocompatible and eco-friendly. It has been used singularly and in combination with other nanomaterials to optimize biosensor design. The attachment of peptides and proteins to nanocrystalline cellulose and their proven retention of activity provide a route to bioactive conjugates useful in designs for point of care biosensors. Elastase is a biomarker for a number of inflammatory diseases including chronic wounds, and its rapid sensitive detection with a facile approach to sensing is of interest. An increased interest in the use of elastase sensors for point of care diagnosis is resulting in a variety of approaches to elsastase sensors utilizing different detection technologies. Here elastase substrate peptide-celluose conjugates synthesized as colorimetric and fluorescent sensors on cotton cellulose nanocrystals are compared. The structure of the sensor peptide-nanocellulose crystals when modeled with computational crystal structure parameters demonstrates the spatio-stoichiometric features of the nanocrystalline surface that allows ligand to active site protease interacttion. An understanding of the structure/function relations of enzyme and conjugate substrate of the peptides covalently attached to nancellulose has implications for enhancing the biomolecular transducer. The potential applications of both fluorescent and colorimetric detection to markers like elastase using peptide cotton cellulose nanocrystals as a transducer surface to model point of care biosensors for protease detection are discussed.

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“…We determined K m and V max for the elastase activity in the HBSS-soluble extracts. K m was 1.1 mM, which compares with a reported K m of 6.2 mM for porcine pancreatic elastase and 0.14 mM for human leukocyte elastase (Castillo et al, 1979). Whether the elastase that degrades endostatin is the elastase secreted into the GI tract remains unclear at present.…”

Section: Discussionmentioning

confidence: 59%

Endostatin expression in pancreatic tissue is modulated by elastase

2004

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Pancreatic tumours are scirrhous, avascular tumours, suggesting that they may produce angiogenesis inhibitors that suppress the growth of the vasculature to the tumour and metastases. We have sought evidence for the angiogenesis inhibitor, endostatin, in normal and cancerous pancreatic tissue. Using Western blotting, we found mature 20 kDa endostatin in cancer tissue but not in normal tissue. Several endostatin-related peptides of higher mol wt were present in both tissues. Extracts from normal tissue were able to degrade exogenous endostatin, whereas extracts from cancer were without effect. Although the exocrine pancreas secretes inactive proenzymes of trypsin, chymotrypsin and elastase, their possible role in this degradation was examined. The trypsin/ chymotrypsin inhibitor, Glycine max, did not prevent the degradation of endostatin by normal pancreatic extracts but elastatinal, a specific inhibitor of elastase, reduced the rate of degradation. Extracts of pancreatic tumours did not express any detectable elastase activity, but an elastase (K m 1.1 mM) was expressed by extracts of normal pancreas. We conclude that endostatin is present and stable in pancreatic cancer tissues, which may explain their avascular nature, but that normal pancreatic tissue expresses enzymes, including elastase, which rapidly degrade endostatin. The stability of endostatin may have implications for its therapeutic use.

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Sensitive substrates for human leukocyte and porcine pancreatic elastase: A study of the merits of various chromophoric and fluorogenic leaving groups in assays for serine proteases

Cited by 245 publications

References 28 publications

IL-23-dependent IL-17 production is essential in neutrophil recruitment and activity in mouse lung defense against respiratory Mycoplasma pneumoniae infection

IL-23-dependent IL-17 production is essential in neutrophil recruitment and activity in mouse lung defense against respiratory Mycoplasma pneumoniae infection

Nanocellulose-Based Biosensors: Design, Preparation, and Activity of Peptide-Linked Cotton Cellulose Nanocrystals Having Fluorimetric and Colorimetric Elastase Detection Sensitivity

Endostatin expression in pancreatic tissue is modulated by elastase

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