A proteolytic fragment of Trypanosoma cruzi trans-sialidase lacking the carboxyl-terminal domain is active, monomeric, and generates antibodies that inhibit enzymatic activity.

Schenkman, Sérgio; Chaves, Luciana Botelho; Carvalho, Lain Carlos Pontes de; Eichinger, Daniel

doi:10.1016/s0021-9258(17)37146-6

Cited by 63 publications

(22 citation statements)

References 30 publications

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“…Ligands. TS used in this study was a His-tagged 70 kDa recombinant protein truncated to remove C-terminal repeats but retaining the catalytic N-terminal domain of the enzyme 33 that contains several antigenic determinants located in solventexposed areas. 37 CrP was obtained essentially homogeneous, starting from crude T. cruzi epimastigote Y strain extracts, in one step, by affinity chromatography.…”

Section: ■ Methodsmentioning

confidence: 99%

“…Although previous studies have reported that the catalytic domain of TS can be used for the diagnosis of chronic CD, 29 the fact that a TS-based SPR immunosensor cannot distinguish between sera from T. cruzi infected patients and control individuals (Figure 2B) can be related to the use of higher serum dilutions in comparison to ELISA, which hinders the possibility of detecting the specific low titer antibodies. 33,34 The possibility to enhance the detection of the specific anti-T. cruzi antibodies in Au-MPA-CrP and Au-MPA-TS sensor platforms by minimizing the impact of non-specific adsorption of serum proteins to the sensor surface still exists. In Figure 3, the relative Δθ SPR (Δθ POS /Δθ NEG ) corresponding to the interaction of positive and negative serum dilutions with Au-MPA-CrP and Au-MPA-TS after the blockage of the nonreactive sites with 1% BSA or 1 M ethanolamine is presented.…”

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confidence: 99%

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Trypanosoma cruzi Virulence Factors for the Diagnosis of Chagas’ Disease

et al. 2019

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trans-Sialidase and cruzipain are important virulence factors from Trypanosoma cruzi, the etiological agent of Chagas disease, that have highly antigenic domains in their structure and were reported as potential tools for diagnosis of the illness. The aim of the present study is to assess the possibility of using cruzipain and the catalytic domain of trans-sialidase in a Surface Plasmon Resonance-based immunosensor for the diagnosis of chronic Chagas disease. Immunoassays carried out with canine sera verified that cruzipain allows the detection of anti-Trypanosoma cruzi antibodies whereas recombinant trans-sialidase did not yield specific detections, due to the high dilutions of serum used in the immunoassays that hinder the possibility to sense the specific low titer antibodies. The developed cruzipain-based biosensor, whose price per assay is comparable to a commercial enzyme-linked immunosorbent assay (ELISA), was successfully applied for the rapid quantification of specific antibodies against Trypanosoma cruzi in fresh human sera showing an excellent agreement with ELISA.

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Section: ■ Methodsmentioning

confidence: 99%

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confidence: 99%

Trypanosoma cruzi Virulence Factors for the Diagnosis of Chagas’ Disease

et al. 2019

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“…The trans-sialidase used in this study was a His-tagged 70 kDa recombinant material truncated to remove C-terminal repeats but retaining the catalytic N-terminal domain of the enzyme. 22 The preparation of the compound libraries screened in this study § Elsewhere we have exploited these observations in the design of novel TcTS acceptor analogue inhibitors 11e and glycomacrocycle substrates 32 for TcTS based on triazole-linked 6-disubstituted b-galactosides.…”

Section: Methodsmentioning

confidence: 99%

Probing the acceptor substrate binding site of Trypanosoma cruzi trans-sialidase with systematically modified substrates and glycoside libraries

et al. 2011

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“…Smith and Eichinger reported the expression and characterisation of Trypanosoma cruzi TS (TcruTS)/ Trypanosoma rangeli sialidase (TranSA) hybrid proteins, exhibiting different Sia transfer and sialidase activities [16]. They found that the C-terminal Fn3 domain (fibronectin type III), named according to its structural relation to fibronectin type III, is not only required for expression of enzymatically active TcruTS and TranSA [17][18][19], but also influences the overall Sia transfer and sialidase activities [16]. Amino acid sequence alignments of a well characterised sialidase from Macromonospora viridifaciens [20] with TcruTS revealed that R572 and E578, which are known to be essential for galactose binding of M. viridifaciens sialidase [20], are well conserved in TcruTS and TranSA [16].…”

Section: Introductionmentioning

confidence: 99%

Cooperativity of catalytic and lectin-like domain of Trypanosoma congolense trans-sialidase modulates its catalytic activity

et al. 2022

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Trans-sialidases (TS) represent a multi-gene family of unusual enzymes, which catalyse the transfer of terminal sialic acids (Sia) from sialoglycoconjugates to terminal galactose or N-acetylgalactosamine residues of oligosaccharides without the requirement of CMP-Neu5Ac, the activated Sia used by typical sialyltransferases. Enzymes comprise a N-terminal catalytic domain (CD) followed by a lectin-like domain (LD). Most work on trypanosomal TS has been done on enzymatic activities focusing on the CD of TS from Trypanosoma cruzi (causing Chagas disease in Latin America), subspecies of Trypanosoma brucei, (causing human sleeping sickness in Africa) and Trypanosoma congolense (causing African Animal Trypanosomosis in livestock). Previously, we demonstrated that T. congolense TS (TconTS)-LD binds to several carbohydrates, such as 1,4-β-mannotriose. In this study we investigated the influence of TconTS3-LD on Sia transfer efficiency of TconTS1a-CD by swapping domains. in silico analysis on structure models of TconTS enzymes revealed the potential of domain swaps between TconTS1a and TconTS3 without structural disruptions of the enzymes overall topologies. Recombinant domain swapped TconTS1a/TS3 showed clear Sia transfer activity, when using fetuin and lactose as Sia donor and acceptor substrates, respectively. While Sia transfer activity remained unchanged from the level of TconTS1a, hydrolytic release of free Neu5Ac as a side product was suppressed resulting in increased transfer efficiency. Presence of 1,4-β-mannotriose during TS reactions modulates enzyme activities enhancing transfer efficiency possibly due to occupation of the binding site in TconTS1a-LD. Interestingly this effect was in the same range as that observed when swapping TconTS1a-CD and TconTS3-LD. In summary, this study demonstrate the proof-of-principle for swapping CDs and LDs of TconTS and that TconTS3-LD influences enzymatic activity of TconTS1a-CD providing evidence that LDs play pivotal roles in modulating activities and biological functions of TconTS and possibly other TS.

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A proteolytic fragment of Trypanosoma cruzi trans-sialidase lacking the carboxyl-terminal domain is active, monomeric, and generates antibodies that inhibit enzymatic activity.

Cited by 63 publications

References 30 publications

Trypanosoma cruzi Virulence Factors for the Diagnosis of Chagas’ Disease

Trypanosoma cruzi Virulence Factors for the Diagnosis of Chagas’ Disease

Probing the acceptor substrate binding site of Trypanosoma cruzi trans-sialidase with systematically modified substrates and glycoside libraries

Cooperativity of catalytic and lectin-like domain of Trypanosoma congolense trans-sialidase modulates its catalytic activity

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