Stoichiometry and heterogeneity of the pro-region chain in tetrameric human cathepsin C

Cigić, Blaž; Križaj, Igor; Kralj, Bogdan; Türk, Vito; Pain, Roger H.

doi:10.1016/s0167-4838(97)00173-8

Cited by 37 publications

(45 citation statements)

References 13 publications

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“…The production of mature, active cathepsin C requires endoproteolytic processing of the proenzyme to remove the internal proregion (44). To determine whether DPAP1 is similarly processed, DPAP1 polypeptides were immunoprecipitated from 35 S-labeled trophozoites (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In addition, three polypeptides with apparent molecular masses of 18 -21 kDa were immunoprecipitated. By analogy with cathepsin C, these polypeptides were presumed to correspond to mature, active DPAP1 generated by excision of the prodomain and a single cleavage within the catalytic region (44). To confirm this, DPAP1 activity was partially purified from trophozoite extract by column chromatography and was subjected to immunoblotting with two DPAP1-specific antibodies (Fig.…”

Section: Resultsmentioning

confidence: 99%

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A Plasmodium falciparum Dipeptidyl Aminopeptidase I Participates in Vacuolar Hemoglobin Degradation

Klemba

Gluzman

Goldberg

2004

Journal of Biological Chemistry

171

174

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Intraerythrocytic growth of the human malaria parasite Plasmodium falciparum requires the catabolism of large amounts of host cell hemoglobin. Endoproteolytic digestion of hemoglobin to short oligopeptides occurs in an acidic organelle called the food vacuole. How amino acids are generated from these peptides is not well understood. To gain insight into this process, we have studied a plasmodial ortholog of the lysosomal exopeptidase cathepsin C. The plasmodial enzyme dipeptidyl aminopeptidase 1 (DPAP1) was enriched from parasite extract by two different approaches and was shown to possess hydrolytic activity against fluorogenic dipeptide substrates. To localize DPAP1 we created a transgenic parasite line expressing a chromosomally encoded DPAP1-green fluorescent protein fusion. Green fluorescent protein fluorescence was observed in the food vacuole of live transgenic parasites, and anti-DPAP1 antibody labeled the food vacuole in parasite cryosections. Together these data implicate DPAP1 in the generation of dipeptides from hemoglobin-derived oligopeptides. To assess the significance of DPAP1, we attempted to ablate DPAP1 activity from blood stage parasites by truncating the chromosomal DPAP1-coding sequence. The inability to disrupt the coding sequence indicates that DPAP1 is important for asexual proliferation. The proenzyme form of DPAP1 was found to accumulate in the parasitophorous vacuole of mature parasites. This observation suggests a trafficking route for DPAP1 through the parasitophorous vacuole to the food vacuole.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

A Plasmodium falciparum Dipeptidyl Aminopeptidase I Participates in Vacuolar Hemoglobin Degradation

Klemba

Gluzman

Goldberg

2004

Journal of Biological Chemistry

171

174

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“…The indicated sites of processing are according to (i) the N-terminal sequences of the peptide chains of native rat DPPI determined by amino acid sequencing (9) (cathepsin J is identical to cathepsin C), and (ii) the homologue sites of native human DPPI (8). In human DPPI, 87 amino acid residues of the C-terminus of the propeptide region is cleaved off and lost upon maturation, and an additional cleavage site at approximately 60 residues from the N-terminus of the propeptide is found in some preparations (7). The polyhistidine tag sequence of HT-rDPPI is fused to the C-terminus of the light chain (shown in bold).…”

Section: Expression Of Rdppi In Insect Cellsmentioning

confidence: 99%

“…Unlike the other members of the papain family, mature DPPI consists of four subunits, each composed of a substantial part of the propeptide and a heavy and light chain derived from cleavage of the catalytic region. Both the propeptide and the heavy chain are glycosylated (7)(8)(9).…”

mentioning

confidence: 99%

Active Recombinant Rat Dipeptidyl Aminopeptidase I (Cathepsin C) Produced Using the Baculovirus Expression System

Lauritzen¹,

Pedersen²,

Madsen³

et al. 1998

Protein Expression and Purification

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“…However, structural modifications of the basic papain framework are present in four members of this family, and these render cathepsins H, C, B and X into an aminopeptidase, dipeptidylpeptidase, peptidyldipeptidase and carboxypeptidase, which remove single residues or dipeptides from the N-or C-terminus respectively. In the case of cathepsins H and C, regions of the propeptide remaining following processing of the proenzyme are responsible for the ability of these enzymes to remove one or two residues respectively from the Nterminus [33,34]. In the case of cathepsins X and B, an additional peptide loop has been grafted on to the basic structure.…”

Section: Discussionmentioning

confidence: 99%

S′2 substrate specificity and the role of His110 and His111 in the exopeptidase activity of human cathepsin B

Krupa

Hasnain

Nägler

et al. 2002

Biochemical Journal

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The ability of the lysosomal cysteine protease cathepsin B to function as a peptidyldipeptidase (removing C-terminal dipeptides) has been attributed to the presence of two histidine residues (His110 and His111) present in the occluding loop, an extra peptide segment located in the primed side of the active-site cleft. Whereas His111 is unpaired, His110 is present as an ion pair with Asp22 on the main body of the protease. This ion pair appears to act as a latch to hold the loop in a closed position. The exopeptidase activity of cathepsin B, examined using quenched fluorescence substrates, was shown to have a 20-fold preference for aromatic side chains in the P′3 position relative to glutamic acid as the least favourable residue. Site-directed mutagenesis demonstrated that His111 makes a positive 10-fold contribution to the exopeptidase activity, whereas His110 is critical for this action with the Asp22—His110 ion pair stabilizing the electrostatic interaction by a maximum of 13.9kJ/mol (3.3kcal/mol). These studies showed that cathepsin B is optimized to act as an exopeptidase, cleaving dipeptides from protein substrates in a successive manner, because of its relaxed specificity in P′3 and its other subsites.

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Stoichiometry and heterogeneity of the pro-region chain in tetrameric human cathepsin C

Cited by 37 publications

References 13 publications

A Plasmodium falciparum Dipeptidyl Aminopeptidase I Participates in Vacuolar Hemoglobin Degradation

A Plasmodium falciparum Dipeptidyl Aminopeptidase I Participates in Vacuolar Hemoglobin Degradation

Active Recombinant Rat Dipeptidyl Aminopeptidase I (Cathepsin C) Produced Using the Baculovirus Expression System

S′2 substrate specificity and the role of His110 and His111 in the exopeptidase activity of human cathepsin B

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