Mapping the extended substrate binding site of cathepsin G and human leukocyte elastase. Studies with peptide substrates related to the alpha 1-protease inhibitor reactive site.

Nakajima, Kiichiro; Powers, James C.; Ashe, Bonnie M.; Zimmerman, M.D.

doi:10.1016/s0021-9258(18)50690-6

Cited by 577 publications

(94 citation statements)

References 18 publications

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“…Next, we addressed the question whether human neutrophil granulocytes (polymorphonuclear neutrophils,P MN) release sufficient amounts of neutrophil elastase upon stimulation for the cleavage of the linker in 6.A ccording to previous investigations,n eutrophil granulocytes contain an average amount of 1.59 pg neutrophil elastase per cell, [22] which corresponds to 1.59 mgmL À1 for 1 10 6 cells mL À1 ,o r to 100 mU mL À1 . [23] Thus,i fa bout 10 %o ft he NE was released or functionally active on the surface of the cells upon stimulation of the granulocytes,s ufficient activity should be available for am easureable cleavage of the conjugates.T o probe this experimentally,P MN were isolated from fresh whole blood, taking into account that the cells are very sensitive and readily activated by different stimuli, including their physical handling. We applied am ild, so-called untouched isolation procedure that removes contaminating cells out of suspension by iron-conjugated antibodies and magnets.…”

Section: Methodsmentioning

confidence: 99%

Selective Bacterial Targeting and Infection‐Triggered Release of Antibiotic Colistin Conjugates

et al. 2021

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In order to render potent, but toxic antibiotics more selective,w eh ave explored an ovel conjugation strategy that includes drug accumulation followed by infection-triggered release of the drug. Bacterial targeting was achieved using am odified fragment of the human antimicrobial peptide ubiquicidin, as demonstrated by fluorophore-tagged variants.To limit the release of the effector colistin only to infectionrelated situations,w ei ntroduced al inker that was cleaved by neutrophil elastase (NE), an enzyme secreted by neutrophil granulocytes at infection sites.The linker carried an optimized sequence of amino acids that was required to assure sufficient cleavage efficiency.T he antibacterial activity of five regioisomeric conjugates prepared by total synthesis was masked, but was released upon exposure to recombinant NE when the linker was attached to amino acids at the 1-or the 3-position of colistin. Ap roof-of-concept was achieved in co-cultures of primary human neutrophils and Escherichia coli that induced the secretion of NE, the release of free colistin, and an antibacterial efficacy that was equal to that of free colistin.

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Section: Methodsmentioning

confidence: 99%

Selective Bacterial Targeting and Infection‐Triggered Release of Antibiotic Colistin Conjugates

et al. 2021

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“…Neutrophil elastase was measured with established in-house assays consisting of elastase-α 1 -antitrypsin soluble product (bound elastase) by sandwich enzyme-linked immunoabsorbent assay 11 using sheep antihuman elastase (Serotec, Oxford, UK), and rabbit antihuman α 1 -antitrypsin (Dako, Bucks, UK), and horseradish peroxidase-labeled swine antirabbit immunoglobulin (affinity purified; Dako). Free elastase was measured by colorimetric assay 12 using MeO-Suc-Ala-Ala-Pro-Val-pNa elastase substrate (Sigma, Dorset, UK), with human leucocyte elastase (Calbiochem, Nottingham, UK) as the standard.…”

Section: Methodsmentioning

confidence: 99%

Paradoxical Neutrophil Elastase Release in Endovascular Abdominal Aortic Aneurysm Repair

Rowlands

Homer‐Vanniasinkam

2007

Vasc Endovascular Surg

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Endovascular repair of abdominal aortic aneurysm potentially avoids problems associated with prolonged aortic cross-clamping that occurs with open repair, but it appears to have its own biologic consequences, which may relate to neutrophil elastase release. Blood samples of consecutive patients undergoing open or endovascular abdominal aneurysm repair were analyzed for neutrophil elastase/alpha(1)-antitrypsin complex and free elastase. Free elastase rose from baseline and fell quickly in open repair patients, returning to baseline by 144 hours. In the endovascular repair group, it continued to increase for up to 144 hours. Bound elastase increased to 24 hours, returning to baseline in endovascular repair patients by 72 hours, but remaining elevated in open repair patients at 144 hours. Open repair patients showed raised elastase/alpha(1)-antitrypsin complex and initial raised free elastase levels. High free elastase levels in endovascular repair patients may reflect less bound elastase and may paradoxically lead to a prolonged inflammatory postoperative response.

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“…TNOB, NFJC and NLEE in concentrations of 1.0, 0.5, and 0.1 mg/mL were evaluated for their inhibitory effects on cathepsin G (Cat-G) enzymes in in-vitro bioassays, in duplicates, according to an established protocol (Nakajima et al, 1979).…”

Section: Cathepsin G Assaymentioning

confidence: 99%

Sunburn (fohia) Healing Effects of Noni: Is it a Mechanism Involving Its Inhibitory Effects on MMP, COX-2 and Cat-G Enzymes?

Palu¹

2012

J. App. Pharm. Sci.

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The aim of the present investigation was to evaluate whether extracts and preparations of noni fruit and leaf, a Polynesian traditional herbal medicine, inhibit MMP, COX-2 and Cat-G enzymes in vitro, as its mechanism of action for healing sun-burn known as fohia in Tonga. Noni leaf ethanolic extract (NLEE) inhibited MMP-1, -2, -3, and -9 enzymes concentration dependently with 0.517, 0.234, 0.184, and 0.302 mg/mL IC 50 , respectively. Noni fruit juice concentrates (NFJC) in 1 and 5 mg/mL concentrations, inhibited MMP-12 enzymes by 102, and 99%, respectively. NFJC and NLEE inhibited Cat-G enzymes concentration-dependently with 0.125, <0.1, and 0.41 mg/mL IC 50 , respectively. Noni fruit juice fractions 4 and 6 inhibited COX-2 and Cat-G enzymes by 85 and 89%, and 89 and 78%, respectively. Additionally, the noni fruit puree and noni leaf has 1.91 mg/g and 5.77 mg/g of ursolic acid, respectively. NFJC, and NLEE inhibitory effects on MMP, COX-2 and Cat-G enzymes might help explain the traditional usage of the noni fruits and leaves for treatment of fohia skin as alluded to by Polynesian traditional healers. These results warrant further studies into the skin health benefits of noni fruit and leaf to further assess their efficacies and dosages in human subjects suffering from photoaging.

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Mapping the extended substrate binding site of cathepsin G and human leukocyte elastase. Studies with peptide substrates related to the alpha 1-protease inhibitor reactive site.

Cited by 577 publications

References 18 publications

Selective Bacterial Targeting and Infection‐Triggered Release of Antibiotic Colistin Conjugates

Selective Bacterial Targeting and Infection‐Triggered Release of Antibiotic Colistin Conjugates

Paradoxical Neutrophil Elastase Release in Endovascular Abdominal Aortic Aneurysm Repair

Sunburn (fohia) Healing Effects of Noni: Is it a Mechanism Involving Its Inhibitory Effects on MMP, COX-2 and Cat-G Enzymes?

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