Role of the FAD-dependent polyamine oxidase in the selective formation of <i>N</i>1,<i>N</i>8-bis(γ- glutamyl)spermidine protein cross-links

Lentini, Alessandro; Mattioli, Palma; Provenzano, Bruno; Abbruzzese, Alberto; Caraglia, Michele; Beninati, Simone

doi:10.1042/bst0350396

Cited by 5 publications

(6 citation statements)

References 25 publications

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“…Further studies are clearly needed in order to assess the role of APAO and SMO in polyamine analogue-mediated drug response. Large versatility in their substrate properties and flexible cleavage of their substrates could implicate that SMO and APAO may have other natural substrates in addition to natural polyamines (Bacchi et al 2009; Lentini et al 2007). It should be noted that the studied analogues resembled SPM (3-4-3) carbon backbone.…”

Section: Discussionmentioning

confidence: 99%

Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases

et al. 2009

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SUMMARYN-alkylated polyamine analogues have potential as anticancer and antiparasitic drugs. However, their metabolism in the host has remained incompletely defined thus potentially limiting their utility. Here, we have studied the degradation of three different spermine analogues N, N′-bis-(3-ethylaminopropyl)butane-1,4-diamine (DESPM), N-(3-benzyl-aminopropyl)-N'-(3-ethylaminopropyl)butane-1,4-diamine (BnEtSPM) and N,N′-bis-(3-benzylaminopropyl)butane-1,4-diamine (DBSPM) and related mono-alkylated derivatives as substrates of recombinant human polyamine oxidase (APAO) and spermine oxidase (SMO). APAO and SMO metabolized DESPM to EtSPD (K m(APAO) =10μM, k cat(APAO) =1.1s −1 and K m(SMO) =28μM, k cat(SMO) =0.8s −1 , respectively), metabolized BnEtSPM to EtSPD (K m(APAO) =0.9 μM, k cat(APAO) =1.1s −1 and K m(SMO) =51μM, k cat(SMO) =0.4s −1 , respectively), and metabolized DBSPM to BnSPD (K m(APAO) =5.4μM, k cat(APAO) = 2.0s −1 and K m(SMO) =33μM, k cat(SMO) =0.3s −1 , respectively). Interestingly, mono-alkylated spermine derivatives were metabolized by APAO and SMO to SPD (EtSPM K m(APAO) =16μM, k cat(APAO) =1.5s −1 ; K m(SMO) =25μM, k cat(SMO) =8.2s −1 ; BnSPM K m(APAO) =6.0μM, k cat(APAO) =2.8s −1 ; K m(SMO) =19μM, k cat(SMO) =0.8s −1 , respectively). Surprisingly, E t S P D ( K m(APAO) =37μM, k cat(APAO) =0.1s −1 ; K m(SMO) =48μM, k cat(SMO) =0.05s −1 ) and BnSPD (K m(APAO) =2.5μM, k cat(APAO) =3.5s −1 ; K m(SMO) =60μM, k cat(SMO) =0.54s −1 ) were metabolized to SPD by both the oxidases. Furthermore, we studied the degradation of DESPM, BnEtSPM or DBSPM in the DU145 prostate carcinoma cell line. The same major metabolites EtSPD and/or BnSPD were detected both in the culture medium and intracellularly after 48 hours of culture. Moreover, EtSPM and BnSPM were detected from cell samples. Present data shows that inducible SMO parallel with APAO could play an important role in polyamine based drug action, i.e. degradation of parent drug and its metabolites, having significant impact on efficiency of these drugs, and hence for the development of novel N-alkylated polyamine analogues.

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Section: Discussionmentioning

confidence: 99%

Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases

et al. 2009

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“…Some experimental strategies have made it possible to evaluate the possibility of the activity of an enzyme, the FAD-PAO, involved in the catabolism of polyamines and in particular of the N 1 - mono (γ-glutamyl)SPD derivative. Indeed, as already reported, under these experimental conditions, the enzyme FAD-PAO is capable of catalyzing the in vitro degradation of peptide-bound N 1 - mono (γ-glutamyl)SPD and N 8 - mono (γ-glutamyl)SPM [ 15 ]. This catabolic process produces free polyamines and peptide-bound γ-glutamyl-3-aminopropionaldehyde.…”

Section: Discussionmentioning

confidence: 70%

“…The observed drastic reduction of the N 1 - mono (γ-glutamyl)SPD levels observed in the lens incubated in the presence of SPD as opposed to N 8 - mono (γ-glutamyl)-SPD levels ( Table 1 ), led to the assumption of a possible degradation mechanism of the N 1 - mono derivative. Based on a previous experience [ 15 ] that showed that the N 1 - mono derivative of SPD is an excellent substrate of the FAD-PAO, the possible role of this enzyme in this phenomenon was investigated. Since rabbit lens FAD-PAO activity, in the presence of SPD was found increased ( Figure 4 ) lenses were cultured with a FAD-PAO inhibitor, the MDL 72527 [ 22 , 23 ], exogenous SPD and Ca 2+ as reported.…”

Section: Resultsmentioning

confidence: 99%

“…In this condition, it was evidenced by the presence of an increased amount of both the mono (γ-glutamyl)SPD derivatives. Interestingly, a degradative pathway of γ-glutamyl-polyamines mediated by flavin adenine dinucleotide FAD-PAO has been previously described [ 15 ]. In the present investigation, pieces of evidence were collected showing that the post-translational modification of β H-crystallins, catalyzed by TG2 is modulated by the levels of lens SPD and regulated by the activity of FAD-PAO, through the degradation of one of the two TG2-formed mono -SPD derivatives, essential for polymerization of β H-crystallins.…”

Section: Introductionmentioning

confidence: 99%

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Polyamine Oxidase Is Involved in Spermidine Reduction of Transglutaminase Type 2-Catalyzed βH-Crystallins Polymerization in Calcium-Induced Experimental Cataract

Mischiati

Feriotto

Tabolacci

et al. 2020

IJMS

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In an in vitro Ca2+-induced cataract model, the progression of opacification is paralleled by a rapid decrease of the endogenous levels of spermidine (SPD) and an increase of transglutaminase type 2 (TG2, EC 2.3.2.13)-catalyzed lens crystallins cross-linking by protein-bound N1-N8-bis(γ-glutamyl) SPD. This pattern was reversed adding exogenous SPD to the incubation resulting in a delayed loss of transparency of the rabbit lens. The present report shows evidence on the main incorporation of SPD by the catalytic activity of TG2, toward βH-crystallins and in particular to the βB2- and mostly in βB3-crystallins. The increase of endogenous SPD in the cultured rabbit lens showed the activation of a flavin adenine dinucleotide (FAD)-dependent polyamine oxidases (PAO EC 1.5.3.11). As it is known that FAD-PAO degrades the N8-terminal reactive portion of N1-mono(γ-glutamyl) SPD, the protein-bound N8-mono(γ-glutamyl) SPD was found the mainly available derivative for the potential formation of βB3-crystallins cross-links by protein-bound N1-N8-bis(γ-glutamyl)SPD. In conclusion, FAD-PAO degradation of the N8-terminal reactive residue of the crystallins bound N1-mono(γ-glutamyl)SPD together with the increased concentration of exogenous SPD, leading to saturation of glutamine residues on the substrate proteins, drastically reduces N1-N8-bis(γ-glutamyl)SPD crosslinks formation, preventing crystallins polymerization and avoiding rabbit lens opacification. The ability of SPD and MDL 72527 to modulate the activities of TG2 and FAD-PAO involved in the mechanism of lens opacification suggests a potential strategy for the prevention of senile cataract.

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“…68 Of note, the covalent modification of basement membrane proteins (laminin, collagen and diverse types of extracellular matrix proteins) by TG may interfere with the adhesiveness and invasiveness of tumor cells, 69,70 thus further supporting the participation of this enzyme in the migration and adhesion processes occurring during the metastatic spread of cancer cells. TG2 catalyzes the incorporation of spermidine into the components of basal membrane, in a process that generally involves FAD-dependent polyamine oxidase (PAO) 71 and that may be one of the cellular mechanisms regulating the preferential formation of a sterically defined bis(γ-glutamyl)spermidine cross-link. In virtue of this fact, it was investigated the effect of this catalytic activity on the adhesion and invasion capability of murine B16-F10 melanoma cells.…”

Section: Transglutaminases and Cancermentioning

confidence: 99%

Possible roles of transglutaminases in molecular mechanisms responsible for cancer and human neurodegenerative diseases

et al. 2017

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Transglutaminases are a family of Ca2+- dependent enzymes which catalyze posttranslational modifications of proteins. The main activity of these enzymes is the crosslinking of glutaminyl residues of a protein/peptide substrate to lysyl residues of a protein/peptide co-substrate. In addition to lysyl residues, other second nucleophilic co-substrates may include monoamines or polyamines (to form mono- or bi-substituted/ crosslinked adducts) or -OH groups (to form ester linkages). In absence of co-substrates, the nucleophile may be water, resulting in the net deamidation of the glutaminyl residue. Transglutaminase activity has been suggested to be involved in molecular mechanisms responsible for either physiological or pathological processes. In particular, transglutaminase activity has been shown to be responsible for human autoimmune diseases and Celiac Disease is just one of them. Interestingly, cancer and neurodegenerative diseases, such as Alzheimer’s Disease, Parkinson’s Disease, supranuclear palsy and Huntington’s Disease, are characterized in part by aberrant transglutaminase activity and by increased cross-linked proteins in affected tissues. This review describes the possible molecular mechanisms by which these enzymes could be responsible for such diseases and the possible use of transglutaminase inhibitors for patients with diseases characterized by aberrant transglutaminase activity.

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Role of the FAD-dependent polyamine oxidase in the selective formation of N1,N8-bis(γ- glutamyl)spermidine protein cross-links

Cited by 5 publications

References 25 publications

Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases

Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases

Polyamine Oxidase Is Involved in Spermidine Reduction of Transglutaminase Type 2-Catalyzed βH-Crystallins Polymerization in Calcium-Induced Experimental Cataract

Possible roles of transglutaminases in molecular mechanisms responsible for cancer and human neurodegenerative diseases

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