Serine protease inhibitors suppress pancreatic endogenous proteases and modulate bacterial neutral proteases

Nduaguibe, Chikodili C.; Bentsi-Barnes, K.; Mullen, Yoko; Kandeel, Fouad; Al-Abdullah, Ismail H.

doi:10.4161/isl.2.3.11714

Cited by 9 publications

(5 citation statements)

References 41 publications

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“…These proteases could be activated during the digestion process and thus may influence islet quality and yields363738. However, our results showed that our peptide substrate cannot be cleaved by the three predominantly endogenous pancreatic proteases using similar conditions for evaluating microbial enzymes.…”

Section: Discussionmentioning

confidence: 84%

Fluorogenic Peptide Substrate for Quantification of Bacterial Enzyme Activities

Al-Abdullah

Bagramyan

Bilbao

et al. 2017

Sci Rep

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A novel peptide substrate (A G G P L G P P G P G G) was developed for quantifying the activities of bacterial enzymes using a highly sensitive Fluorescence Resonance Energy Transfer (FRET) based assay. The peptide substrate was cleaved by collagenase class I, II, Liberase MTF C/T, collagenase NB1, and thermolysin/neutral protease, which was significantly enhanced in the presence of CaCl2. However, the activities of these enzymes were significantly decreased in the presence of ZnSO4 or ZnCl2. Collagenase I, II, Liberase MTF C/T, thermolysin/neutral protease share similar cleavage sites, L↓G and P↓G. However, collagenase NB1 cleaves the peptide substrate at G↓P and P↓L, in addition to P↓G. The enzyme activity is pH dependent, within a range of 6.8 to 7.5, but was significantly diminished at pH 8.0. Interestingly, the peptide substrate was not cleaved by endogenous pancreatic protease such as trypsin, chymotrypsin, and elastase. In conclusion, the novel peptide substrate is collagenase, thermolysin/neutral protease specific and can be applied to quantify enzyme activities from different microbes. Furthermore, the assay can be used for fine-tuning reaction mixtures of various agents to enhance the overall activity of a cocktail of multiple enzymes and achieve optimal organ/tissue digestion, while protecting the integrity of the target cells.

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Section: Discussionmentioning

confidence: 84%

Fluorogenic Peptide Substrate for Quantification of Bacterial Enzyme Activities

Al-Abdullah

Bagramyan

Bilbao

et al. 2017

Sci Rep

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“…The above maximum inhibition rates of native UTI and those for elastase, described below, supported previous reports. 8) 9) 15) UTI and HA-UTI, both treated with actinase E, did not inhibit activities of these proteases. These results supported that the core protein of UTI has protease inhibitory activity toward a broad range of enzymes previously reported by others.…”

Section: Methodsmentioning

confidence: 89%

“…UTI is a key anti-inflammatory response mediator inhibiting the activities of many enzymes that facilitate inflammation. 1) 8) 9) UTI inhibits serine proteases, such as trypsin, chymotrypsin, plasmin, elastase, thrombin, kallikrein, etc , by binding to them at either of the two Kuniz-type domains of its core protein. Based on the protease inhibitory activity, human UTI is used as an anti-inflammatory medicine, for example, in the treatment for acute pancreatitis, 10) or Kawasaki disease, 11) or for inhibiting threatened premature delivery, 12) etc .…”

mentioning

confidence: 99%

A Chondroitin Sulfate Chain of Urinary Trypsin Inhibitor Enhances Protease Inhibitory Activity of the Core Protein

et al. 2020

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Human urinary trypsin inhibitor (UTI) is a proteoglycan composed of one core protein covalently linked to one glycosaminoglycan, which is a low sulfated chondroitin 4-sulfate. It is used as an anti-inflammatory medicine based on the protease inhibitory activity of the core protein. However, functions of the chondroitin sulfate have not been clarified. Recently, we succeeded in remodeling the UTI chondroitin sulfate to hyaluronan to create hyaluronan hybrid UTI, without changing the core protein. Here, we investigated the effect of the remodeled chondroitin sulfate on the activities of serine proteases. Native UTI showed stronger protease inhibitory activity than hyaluronan hybrid UTI or hydrolyzed glycosaminoglycan UTI. Chondroitin 4-sulfate chains with a small peptide derived from the native UTI did not show any protease inhibitory activity. These results suggest that the chondroitin sulfate chain linked covalently to core protein enhances protease inhibitor activity of UTI although the chondroitin sulfate chain itself does not.

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“…Attempts to use Pefabloc during the isolation process instead of organ preservation phase have been made, showing that the Pefabloc can inhibit serine protease activity throughout the tissue digestion process; but, with controversial results on islet yield and function [94–96]. In addition, two research papers recently published by the same group show that when human islets were cultured or perifused with Pefabloc postisolation, insulin secretion of human islets in response to glucose stimulation was compromised, suggesting that the Pefabloc is not suitable for human islet isolation [97, 98]. …”

Section: Mitochondria and Mitoprotection In Islet Isolation And Cumentioning

confidence: 99%

Implication of Mitochondrial Cytoprotection in Human Islet Isolation and Transplantation

Wang

Mendoza-Elias

et al. 2012

Biochemistry Research International

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Islet transplantation is a promising therapy for type 1 diabetes mellitus; however, success rates in achieving both short- and long-term insulin independence are not consistent, due in part to inconsistent islet quality and quantity caused by the complex nature and multistep process of islet isolation and transplantation. Since the introduction of the Edmonton Protocol in 2000, more attention has been placed on preserving mitochondrial function as increasing evidences suggest that impaired mitochondrial integrity can adversely affect clinical outcomes. Some recent studies have demonstrated that it is possible to achieve islet cytoprotection by maintaining mitochondrial function and subsequently to improve islet transplantation outcomes. However, the benefits of mitoprotection in many cases are controversial and the underlying mechanisms are unclear. This article summarizes the recent progress associated with mitochondrial cytoprotection in each step of the islet isolation and transplantation process, as well as islet potency and viability assays based on the measurement of mitochondrial integrity. In addition, we briefly discuss immunosuppression side effects on islet graft function and how transplant site selection affects islet engraftment and clinical outcomes.

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Serine protease inhibitors suppress pancreatic endogenous proteases and modulate bacterial neutral proteases

Cited by 9 publications

References 41 publications

Fluorogenic Peptide Substrate for Quantification of Bacterial Enzyme Activities

Fluorogenic Peptide Substrate for Quantification of Bacterial Enzyme Activities

A Chondroitin Sulfate Chain of Urinary Trypsin Inhibitor Enhances Protease Inhibitory Activity of the Core Protein

Implication of Mitochondrial Cytoprotection in Human Islet Isolation and Transplantation

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