A trypanosome oligopeptidase as a target for the trypanocidal agents pentamidine, diminazene and suramin

Morty, Rory E.; Troeberg, Linda; Pike, Robert N.; Jones, Roy; Nickel, Peter; Lonsdale‐Eccles, John D.; Coetzer, Theresa H.T.

doi:10.1016/s0014-5793(98)00914-4

Cited by 68 publications

(48 citation statements)

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“…Oligopeptidase B (OPB) 2 has been identified in other protozoa, including T. cruzi, Trypanosoma brucei, and Trypanosoma evansi (7-9). These enzymes represent a subfamily of potential chemotherapeutic targets (10) because their inhibitors have been shown to be trypanocidal (4,11,12). OPB of T. cruzi is involved in the invasion of host cells through the generation of an unknown signaling ligand that interacts with host cells in order to recruit lysosomes to their surface (7).…”

mentioning

confidence: 99%

The Oligopeptidase B of Leishmania Regulates Parasite Enolase and Immune Evasion

Swenerton¹,

Zhang²,

Sajid

et al. 2011

Journal of Biological Chemistry

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Proteases are a ubiquitous group of enzymes that play key roles in the life cycle of parasites, in the host-parasite relationship, and in the pathogenesis of parasitic diseases. Furthermore, proteases are targets for the development of new antiparasitic therapy. Protozoan parasites like Leishmania predominantly express Clan CA cysteine proteases for key life cycle functions. It was therefore unexpected to find a high level of serine protease activity expressed by Leishmania donovani. Purification of this activity followed by mass spectrometry identified oligopeptidase B (OPB; Clan SC, family S9A) as the responsible enzyme. This was confirmed by gene knock-out of OPB, which resulted in the disappearance of the detected serine protease activity of Leishmania extracts. To delineate the specific role of OPB in parasite physiology, proteomic analysis was carried out on OPB(؊/؊) versus wild type parasites. Four protein species were significantly elevated in OPB(؊/؊) parasites, and all four were identified by mass spectrometry as enolase. This increased enolase was enzymatically inactive and associated with the parasite membrane. Aside from its classic role in carbohydrate metabolism, enolase was recently found to localize to membranes, where it binds host plasminogen and functions as a virulence factor for several pathogens. As expected, there was a striking alteration in macrophage responses to Leishmania when OPB was deleted. Whereas wild type parasites elicited little, if any, response from infected macrophages, OPB(؊/؊) parasites induced a massive up-regulation in gene transcription. Additionally, these OPB(؊/؊) parasites displayed decreased virulence in the murine footpad infection model.Proteases are a ubiquitous group of enzymes functioning in nearly all biological phenomena. In protozoan parasites, proteases play key roles in life cycle transition, host invasion, parasite immune evasion, and the pathogenesis of parasitic diseases (1, 2). At least 50 cysteine proteases and twenty serine proteases are known to exist in the Leishmania genome (3-5).The molecular evolution of proteases shows a striking dichotomy between the predominant Clan CA cysteine proteases of protozoa and primitive metazoa and the dominant trypsin family (S1A) serine proteases of vertebrates (6). It was therefore unexpected to find that Leishmania donovani, Leishmania major, and Leishmania mexicana (but not the related kinetoplastid parasite, Trypanosoma cruzi) express high levels of serine protease activity, relative to cysteine protease activity in parasite extracts.To delineate the character of this serine protease activity, it was purified from Leishmania extracts and identified as Clan SC, family S9A oligopeptidase B by mass spectrometry. Oligopeptidase B (OPB) 2 has been identified in other protozoa, including T. cruzi, Trypanosoma brucei, and Trypanosoma evansi (7-9). These enzymes represent a subfamily of potential chemotherapeutic targets (10) because their inhibitors have been shown to be trypanocidal (4,11,12). OPB of T. cruzi is i...

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confidence: 99%

The Oligopeptidase B of Leishmania Regulates Parasite Enolase and Immune Evasion

Swenerton¹,

Zhang²,

Sajid

et al. 2011

Journal of Biological Chemistry

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“…In protozoa, OPB may be involved in the pathogenesis of several diseases, and it has been identified as a virulence factor and potential drug target. [11][12][13][14][15] OPB was originally found in bacteria and it was purified from E. coli. 9) Although the involvement of OPB in the infection of bacteria was not established, active OPB enzymes may play a role as observed for the pathogenesis of trypanosomes.…”

Section: Discussionmentioning

confidence: 99%

“…In fact, it has been identified as a target of several drugs (pentamidine, diminazene and suramin) used in the treatment of African trypanosomiasis. 12) Administration of irreversible OPB inhibitors to trypanosome-infected mice has been shown to significantly impair disease progression, and irreversible inhibitors of the enzyme exhibit antitrypanosomal activity in vitro and in vivo. 13) The mechanism by which Trypanosoma brucei and T. evanci OPBs cause African sleeping sickness and surra may involve the release of these pathogens from dying parasites into the host bloodstream, where they remains stable and catalytically active and degrade biologically active peptides.…”

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confidence: 99%

Assessment of Substrate Inhibition of Bacterial Oligopeptidase B

Mohamed

Nakajima

Oyama

et al. 2012

Biological & Pharmaceutical Bulletin

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Oligopeptidase B (OPB; EC 3.4.21.83) from 2 Gram-negative bacteria, Stenotrophomonas maltophilia (Stm) and Serratia marcescens (Sem), and the Gram-positive bacterium Rhodococcus erythropolis (Re) were cloned and characterized to clarify their activities and substrate specificities using peptidyl-MCA substrates containing Arg or Lys. The cloned enzymes, Stm, Sem and ReOPBs, in addition to Escherichia coli OPB (EcOPB) were expressed using a pET expression system. Although the Stm and SemOPBs share 45% sequence identity to each other and up to 60% identity with respect to their catalytic domains, their activities towards MCA substrates were quite different. StmOPB is approximately 100-500 times more active than SemOPB and 3-30 times more active than EcOPB. The activity of ReOPB is comparable to that of StmOPB and it shares 40% and 36% identity to StmOPB and SemOPB, respectively. Some features of Stm, Re and EcOPBs are similar to those of previously cloned OPBs, which were also strongly inhibited by substrates, but SemOPB differs from all other OPBs in that it is not inhibited by substrates; even substrates containing double arginine at 35 µM did not inhibit SemOPB. On the other hand, the same substrates at only 5 µM inhibited the activity of the Stm, Re, and EcOPB. This phenomenon was not observed with substrates containing single or double lysine.Key words oligopeptidase B; prolyl oligopeptidase family; substrate inhibition; opportunistic bacteria; substrate specificityThe oligopeptidase B (OPB, EC 3.4.21.83) subfamily represents one of two branches of the prolyl oligopeptidase family of serine peptidases (clan SC, family S9).1) The substrate specificities of the subfamily enzymes are different, with prolyl oligopeptidase exclusively hydrolyzing peptide bonds at the C-terminal to proline residues in peptides, 2,3) while OPB demonstrates a trypsin-like substrate specificity, hydrolyzing peptide bonds on the C-terminal side of basic amino acid residues. Arginine-or lysine-containing peptides of no more than about 30 residues can be hydrolyzed, [4][5][6] since the N-terminal β-propeller domain of the enzymes blocks access of large globular proteins to the catalytic machinery.3) However, it has been demonstrated that OPB can cleave, in addition to low-molecular-mass peptides, several basic proteins in a restricted fashion, including human histones H1, H2A, H3, and H4.7) Peptides with Arg residues in both P1 and P2 are hydrolyzed at a much faster rate by OPB than peptides with only one Arg residue at the P1 site. 7,8) OPB is found in bacteria, plants and trypanosomatid pathogens, where it has been identified as a virulence factor and a potential drug target, since the genes coding for this enzyme are absent from mammals. The exact physiological function(s) and the physiological substrates of the OPB are unknown. Although the OPB was first cloned and characterized from Escherichia coli 4,9) and Moraxella lacunata, 6) bacterial OPBs have received much less attention than their homologues from protozoa. Thus their 3D stru...

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“…In this sense, oligopeptidases B display similar properties to several hormone-processing enzymes, thus host-peptide hormones are candidates for cleavage by the parasite oligopeptidases [222]. In fact, the enzyme has been identified as a target of several trypanocidal drugs, such as suramin [223] and its proper characterization may lead to the design of highly selective inhibitors to be employed as trypanocidal drugs [222].…”

Section: Serine Peptidasesmentioning

confidence: 99%

Biological Roles of Peptidases in Trypanosomatids~!2009-11-26~!2010-02-15~!2010-03-18~!

Vermelho

Branquinha²,

d’Avila-Levy³

et al. 2010

TOPARAJ

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Abstract:In this review, we report the recent developments in the characterization of peptidases and their possible biological functions in the Trypanosomatidae family. The focus will be on peptidases from Trypanosoma cruzi, Leishmania spp., African trypanosomes and plant and insect trypanosomatids. There are numerous events in parasite development where the involvement of peptidases has been established, and they will be approached in the present review. Also in this review we will discuss the central roles have been proposed for peptidases in diverse processes such as virulence, host cell interaction and invasion, catabolism of host proteins, differentiation, cell cycle progression and both stimulation and evasion of host immune responses.

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A trypanosome oligopeptidase as a target for the trypanocidal agents pentamidine, diminazene and suramin

Cited by 68 publications

References 42 publications

The Oligopeptidase B of Leishmania Regulates Parasite Enolase and Immune Evasion

The Oligopeptidase B of Leishmania Regulates Parasite Enolase and Immune Evasion

Assessment of Substrate Inhibition of Bacterial Oligopeptidase B

Biological Roles of Peptidases in Trypanosomatids~!2009-11-26~!2010-02-15~!2010-03-18~!

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