Sulfonyl fluoride serine protease inhibitors inactivate RNK-16 lymphocyte granule proteases and reduce lysis by granule extracts and perforin

Ewoldt, Gerald R.; Winkler, Ulrike; Powers, James C.; Hudig, Dorothy

doi:10.1016/0161-5890(92)90181-v

Cited by 22 publications

(6 citation statements)

References 31 publications

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“…Therefore, replacement of the susceptible ester bond of a polyisoprenylated cysteinyl substrate with a reactive “warhead” [5] such as sulfonyl fluoride might convert a high affinity substrate to a highly effective pseudo-substrate irreversible inhibitor of PMPMEase. This is a well established approach for the development of potent inhibitors of serine hydrolases [22, 23]. The polyisoprenylation, besides increasing affinity, also renders the compounds more selective since polyisoprenylated cysteine ester substrates are resistant to hydrolysis by cholinesterases [2].…”

Section: Discussionmentioning

confidence: 99%

Polyisoprenylation Potentiates the Inhibition of Polyisoprenylated Methylated Protein Methyl Esterase and the Cell Degenerative Effects of Sulfonyl Fluorides

Aguilar¹,

Amissah²,

Duverna³

et al. 2011

CCDT

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The polyisoprenylation pathway incorporates a reversible step that metabolizes polyisoprenylated methylated proteins from the ester to the carboxylate form. Polyisoprenylated protein methyl transferase (PPMTase) catalyses the esterification whereas polyisoprenylated methylated protein methyl esterase (PMPMEase) hydrolyzes them. Significant changes in the balance between the two enzymes may alter polyisoprenylated protein function possibly resulting in disease. Previous studies show that PMPMEase is the serine hydrolase, Sus scrofa carboxylesterase. Its susceptibility to the nonspecific serine hydrolase inhibitor, phenylmethylsulfonyl fluoride (PMSF) paved the way for its use as a prototypical compound to design and synthesize a series of putative high affinity specific inhibitors of PMPMEase. Pseudo first-order kinetics revealed an over 680-fold increase in kobs/[I] values from PMSF (6 M−1s−1), S-phenyl (L-50, 180 M−1s−1), S-benzyl (L-51, 350 M−1s−1), S-trans, trans-farnesyl (L-28, 2000 M−1s−1), to S-trans-geranylated (L-23, 4100 M−1s−1) 2-thioethanesulfonyl fluorides. C10 S-alkyl substitution revealed a kobs/[I] value (1800 M−1s−1) that was 298 times greater than that for PMSF. The compounds induced the degeneration of human neuroblastoma SH-SY5Y cells with EC50 values of 49, 130 and >1000 μM for L-28, L-23 and PMSF, respectively. The increased affinity with the polyisoprenyl derivatization is consistent with the observed substrate specificity and the reported hydrophobic nature of the active site. These results suggest that (1) PMPMEase is a key enzyme for polyisoprenylated protein metabolism, (2) regulation of its activity is essential for maintaining normal cell viability, (3) abnormal activities may be involved in degenerative diseases and cancers and (4) its specific inhibitors may be useful in combating cancers.

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Section: Discussionmentioning

confidence: 99%

Polyisoprenylation Potentiates the Inhibition of Polyisoprenylated Methylated Protein Methyl Esterase and the Cell Degenerative Effects of Sulfonyl Fluorides

Aguilar¹,

Amissah²,

Duverna³

et al. 2011

CCDT

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“…Using Z-Arg-Arg-pNA and BSA as substrates, the effect of Cry41-related toxin on Cathepsin B activity was determined and compared with that of different inhibitors , and activators. As shown in Figure A, all the tested inhibitors exhibited obvious inhibition on Cathepsin B activity using BSA substrates, while different concentrations of Cry41-related toxin obviously enhanced Cathepsin B activity.…”

Section: Resultsmentioning

confidence: 99%

Possible Insecticidal Mechanism of Cry41-Related Toxin against Myzus persicae by Enhancing Cathepsin B Activity

Zhao

Zhang

et al. 2020

J. Agric. Food Chem.

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Cry toxins produced by Bacillus thuringiensis are well known for their high insecticidal activities against Lepidoptera, Diptera, and Coleoptera; however, their activities against Aphididae are very low. Recently, it has been reported that a Cry41-related toxin exhibited moderate activity against the aphid Myzus persicae, and thus, it is highly desirable to uncover its unique mechanism. In this paper, we report that Cathepsin B, calcium-transporting ATPase, and symbiotic bacterial-associated protein ATP-dependent-6-phosphofructokinase were pulled down from the homogenate of M. persicae as unique proteins that possibly bound to Cry41related toxin. Cathepsin B has been reported to cleave and inactivate antiapoptotic proteins and plays a role in caspase-initiated apoptotic cascades. In this study, Cathepsin B was expressed in Escherichia coli and purified, and in vitro interaction between recombinant Cathepsin B and Cry41-related toxin was demonstrated. Interestingly, we found that addition of Cry41-related toxin obviously enhanced Cathepsin B activity. We propose a model for the mechanism of Cry41-related toxin as follows: Cry41-related toxin enters the aphid cells and enhances Cathepsin B activity, resulting in acceleration of apoptosis of aphid cells.

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“…The variation in inhibition observed with LAAO with PMSF is worthy of further comment, and not just for the LAAO but for other enzymes that are not serine proteases. First, PMSF does not inhibit all serine proteases [24,25], and not all enzymes that are inhibited by PMSF are serine proteases, which include LAAO [3,4,5], CoA-independent transacylase [26], diacylglycerol lipase [27], calcium-dependent cysteine proteinase [28] and the factor X activator losac, a hemolin [29]. In summary, while the interaction of PMSF with a serine residue within the enzyme catalytic site is the classic paradigm of inhibition, it is entirely possible that there may be proteins with serine residues outside the catalytic site that could modulate enzymatic activity (e.g., changes in structural conformation).…”

Section: Discussionmentioning

confidence: 99%

Characterization of L-amino Acid Oxidase Derived from Crotalus adamanteus Venom: Procoagulant and Anticoagulant Activities

Nielsen

2019

IJMS

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Snake venom enzymes of the L-amino acid oxidase (LAAO) class are responsible for tissue hemorrhage, edema, and derangement of platelet function. However, what role, if any, these flavoenzymes play in altering plasmatic coagulation have not been well defined. Using coagulation kinetomic analyses (thrombelastograph-based), it was determined that the LAAO derived from Crotalus adamanteus venom displayed a procoagulant activity associated with weak clot strength (no factor XIII activation) similar to thrombin-like enzymes. The procoagulant activity was not modified in the presence of reduced glutathione, demonstrating that the procoagulant activity was likely due to deamination, and not hydrogen peroxide generation by the LAAO. Further, unlike the raw venom of the same species, the purified LAAO was not inhibited by carbon monoxide releasing molecule-2 (CORM-2). Lastly, exposure of the enzyme to phenylmethylsulfonyl fluoride (PMSF) resulted in the LAAO expressing anticoagulant activity, preventing contact activation generated thrombin from forming a clot. In sum, this investigation for the first time characterized the LAAO of a snake venom as both a fibrinogen polymerizing and an anticoagulant enzyme acting via oxidative deamination and not proteolysis as is the case with thrombin-like enzymes (e.g., serine proteases). Using this thrombelastographic approach, future investigation of purified enzymes can define their biochemical nature.

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Sulfonyl fluoride serine protease inhibitors inactivate RNK-16 lymphocyte granule proteases and reduce lysis by granule extracts and perforin

Cited by 22 publications

References 31 publications

Polyisoprenylation Potentiates the Inhibition of Polyisoprenylated Methylated Protein Methyl Esterase and the Cell Degenerative Effects of Sulfonyl Fluorides

Polyisoprenylation Potentiates the Inhibition of Polyisoprenylated Methylated Protein Methyl Esterase and the Cell Degenerative Effects of Sulfonyl Fluorides

Possible Insecticidal Mechanism of Cry41-Related Toxin against Myzus persicae by Enhancing Cathepsin B Activity

Characterization of L-amino Acid Oxidase Derived from Crotalus adamanteus Venom: Procoagulant and Anticoagulant Activities

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