Cloning and Characterization of the Schistosoma japonicum Aspartic Proteinase Involved in Hemoglobin Degradation

Becker, Marion; Harrop, Stephen Anthony; Dalton, John P.; Kalinna, Bernd H.; McManus, Donald P.; Brindley, Paul J.

doi:10.1074/jbc.270.41.24496

Cited by 75 publications

(44 citation statements)

References 24 publications

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“…More specifically, the activity of dipeptidylpeptidases (and aminopeptidases) can be expected to supplement the action of schistosome endoproteases including cathepsins B, D and L in the catabolism of the host-derived hemoglobin [7,8,15,16].…”

Section: Resultsmentioning

confidence: 99%

“…The insert of clone pSjC4 was digested with KpnI and BamHI, separated by agarose-gel electrophoresis, purified by Wizard PCR DNA cleanup resin column (Promega), and used as a probe. The insert (50 ng) was radiolabeled with [A-32 P]dCTP and hybridized to the membrane using hybridization and washing conditions described previously [15]. Autoradiography was performed at Ϫ70°C using Kodak X-AR film and intensifying screens.…”

Section: Methodsmentioning

confidence: 99%

“…Gene fragments were radiolabeled with [A-32 P]dCTP (Amersham) by random priming. Hybridization and washing conditions were as previously described [15]. The pBluescript phagemids in positive clones were excised using R408 helper phage (Promega) and E. coli strain XL1-Blue.…”

Section: Methodsmentioning

confidence: 99%

“…Each of the proteolytic enzymes involved is a potential target for novel, antischistosomal therapies [7Ϫ9]. The proteases include the endoproteases cathepsin B Sm31 [10], cathepsin L [11,12], schistosome legumain [13,14] and cathepsin-D-like aspartic protease [15,16]. It is also likely that exopeptidases, such as dipeptidylpeptidases (DPPs) and aminopeptidases, are involved in the hemoglobin digestion subsequent to the initial cleavages of the protein by endopeptidases [9,17].…”

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confidence: 99%

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Cathepsin C from Schistosoma japonicum

Hola-Jamriska

Tort

Dalton

et al. 1998

European Journal of Biochemistry

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A cDNA encoding preprocathepsin C was isolated from adults of the asian blood fluke Schistosoma japonicum. The deduced amino acid sequence of S. japonicum cathepsin C comprised 458 amino acid residues; 22 NH 2 -terminal residues corresponding to the signal peptide, 199 residues corresponding to the propeptide and 237 COOH-terminal residues corresponding to the mature enzyme region. The amino acid sequence of this preprocathepsin showed 43% and 50% identity to that of human and rat, respectively. The preproenzyme shared only 59% identity with the sequence for a cathepsin C reported from Schistosoma mansoni, differing from it in active-site residues and in its potential N-glycosylation sites. Northern-blot analysis showed that S. japonicum cathepsin C was expressed in greater quantities in female than in male parasites. Phylogenetic analysis utilizing the mature enzyme sequences of S. japonicum and other cathepsin Cs demonstrated that cathepsin Cs and cathepsin Bs shared a common ancestry. The unusually long prosegment observed in cathepsin C from S. japonicum and from other species was compared to that of cathepsin Bs and cathepsin Ls. The extension contained two blocks of residues which were highly conserved among cathepsin Cs. The COOH terminus of the prosegment exhibited a composite of features present in the prosegments of cathepsin Ls and cathepsin Bs. Most significantly, given the common ancestry of cathepsin B and cathepsin C, the prosegment of cathepsin C included ERFNIN-like motifs and other residues more characteristic of non-cathepsin-B-like members of the papain superfamily such as cathepsin L.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Cathepsin C from Schistosoma japonicum

Hola-Jamriska

Tort

Dalton

et al. 1998

European Journal of Biochemistry

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“…The highest sequence similarities were found in three invertebrate aspartyl proteases: ASPP_AEDE, a lysosomal enzyme from Aedes aegypti (mosquito) [16], Acasp a aspartyl protease from the anthropophilic hookworm Ancylostoma caninum (nematode) [17] and Sjpasp from the blood fluke Schistosoma japonicum [18] (plathelminth).…”

Section: Correlation Of N-terminal Sequences With Genomic Datamentioning

confidence: 99%

Aspartyl proteases in Caenorhabditis elegans

Geier

Banaj

Heid

et al. 1999

European Journal of Biochemistry

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Crude homogenates of the nematode Caenorhabditis elegans exhibit maximal proteolytic activity under acidic pH conditions. About 90% of this activity is inhibited by the oligopeptide pepstatin, which specifically inhibits the activity of aspartyl proteases such as pepsin, cathepsins D and E or renin.We have purified enzymes responsible for this proteolytic activity by a single-step affinity chromatography on pepstatin±agarose. Analysis of the purified fraction by 1D SDS gel electrophoresis revealed six bands ranging from 35 to 52 kDa. After electrotransfer to poly(vinylidene difluoride) membranes, all bands were successfully subjected to N-terminal microsequencing.On 2D gels, the purified protein bands split into 19 spots which, after renewed microsequencing, were identified as isoelectric variants of the six proteins already described. The N-termini obtained for these proteins could be correlated to genomic DNA sequences determined in the course of the C. elegans genome sequencing project. All these sequences were predicted to code for expressed proteins as collected in the WORMPEP database. Five of the six coding sequences identified in this study were found to contain the typical active-site consensus sequence of aspartyl proteases and displayed an overall amino acid identity between 25 and 66% as compared to aspartyl proteases from other organisms. In addition to the five aspartyl proteases detected at the protein level, we have identified the coding sequences for seven other enzymes of this protease family by a similarity search in the genomic DNA of C. elegans which has recently been completely sequenced.

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Conformational homogeneity in molecular recognition by proteolytic enzymes

Tyndall

Fairlie

1999

J. Mol. Recognit.

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Crystal structures for several hundred protease-inhibitor complexes have been analysed and their superimpositions have been used to demonstrate a universal relationship between inhibitor/substrate conformation and molecular recognition by all aspartic, serine, cysteine and metallo proteases. Proteases universally recognize an extended beta strand conformation in all their peptidic (and non-peptidic) inhibitors and substrate analogues without significant exceptions. This conformational homogeneity is illustrated here for a subset of 180 protease-inhibitor structures which are displayed as (a) structural overlays of multiple inhibitors for each of eight aspartic, eight serine, six metallo and five cysteine proteases; (b) single inhibitors each bound to different proteases; and (c) Ramachandran plots of peptide or pseudo-peptide dihedral angle pairs which demonstrate beta strands (Phi -54 degrees to -173 degrees, Psi 24 degrees to 174 degrees ) like those normally found paired in proteins as beta sheets. However, unlike beta sheets, alpha and 3(10) helices, beta and gamma turns, where the folded main chain amide components are intramolecularly hydrogen bonded and thus unavailable for interaction with proteins, an inhibitor/substrate in an isolated beta strand conformation provides maximum exposure of its hydrogen bonding donors/acceptors and side chain components to a putative protease receptor. This analysis highlights the advantages of a strand conformation over other elements of secondary structure for protease recognition and may lead to generic strategies for inhibitor design.

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Cloning and Characterization of the Schistosoma japonicum Aspartic Proteinase Involved in Hemoglobin Degradation

Cited by 75 publications

References 24 publications

Cathepsin C from Schistosoma japonicum

Cathepsin C from Schistosoma japonicum

Aspartyl proteases in Caenorhabditis elegans

Conformational homogeneity in molecular recognition by proteolytic enzymes

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