Catalytic Domain Architecture of Metzincin Metalloproteases

Gomis‐Rüth, F. Xavier

doi:10.1074/jbc.r800069200

Cited by 206 publications

(188 citation statements)

References 44 publications

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“…Next in similarity was ulilysin, the structural prototype of the pappalysin family, followed by MMPs, serralysins, and astacins, which are all members of the metzincin clan of MPs (7,8). Fig.…”

Section: Resultsmentioning

confidence: 99%

Structure, function and latency regulation of a bacterial enterotoxin potentially derived from a mammalian adamalysin/ADAM xenolog

Goulas

Arolas

Gomis‐Rüth

2011

Proc. Natl. Acad. Sci. U.S.A.

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Enterotoxigenic Bacteroides fragilis is the most frequent diseasecausing anaerobe in the intestinal tract of humans and livestock and its specific virulence factor is fragilysin, also known as B. fragilis toxin. This is a 21-kDa zinc-dependent metallopeptidase existing in three closely related isoforms that hydrolyze E-cadherin and contribute to secretory diarrhea, and possibly to inflammatory bowel disease and colorectal cancer. Here we studied the function and zymogenic structure of fragilysin-3 and found that its activity is repressed by a ∼170-residue prodomain, which is the largest hitherto structurally characterized for a metallopeptidase. This prodomain plays a role in both the latency and folding stability of the catalytic domain and it has no significant sequence similarity to any known protein. The prodomain adopts a novel fold and inhibits the protease domain via an aspartate-switch mechanism. The catalytic fragilysin-3 moiety is active against several protein substrates and its structure reveals a new family prototype within the metzincin clan of metallopeptidases. It shows high structural similarity despite negligible sequence identity to adamalysins/ADAMs, which have only been described in eukaryotes. Because no similar protein has been found outside enterotoxigenic B. fragilis, our findings support that fragilysins derived from a mammalian adamalysin/ADAM xenolog that was co-opted by B. fragilis through a rare case of horizontal gene transfer from a eukaryotic cell to a bacterial cell. Subsequently, this co-opted peptidase was provided with a unique chaperone and latency maintainer in the time course of evolution to render a robust and dedicated toxin to compromise the intestinal epithelium of mammalian hosts.bacterial endotoxin | human pathogen | zymogen activation

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Section: Resultsmentioning

confidence: 99%

Structure, function and latency regulation of a bacterial enterotoxin potentially derived from a mammalian adamalysin/ADAM xenolog

Goulas

Arolas

Gomis‐Rüth

2011

Proc. Natl. Acad. Sci. U.S.A.

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“…Those are the astacins, ADAMs/adamalysins/reprolysins, serralysins, matrix metalloproteases, snapalysins, leishmanolyisns, and pappalysins. 4 With archaemetzincin from Methanopyrus kandleri, we describe for the first time the crystal structure of a member of the archaemetzincin family. Interestingly, we found in addition to the zinc ion in the active site, a second zinc ion coordinated by the four cysteine residues from the consensus sequence, thus resembling a zinc-finger motif.…”

Section: Introductionmentioning

confidence: 99%

Crystal structure of archaemetzincin amza from Methanopyrus kandleri at 1.5 Å resolution

et al. 2010

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“…The metzincins are a clan of MPs characterized by a consensus sequence responsible for binding of the catalytic zinc ion (CSBZ), HEXXHXX(G/N)XX(H/D), and a conserved methionine-containing turn (Met-turn) (12)(13)(14)(15)(16). This clan can be subdivided into several families, including the MMPs.…”

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Proenzyme Structure and Activation of Astacin Metallopeptidase

Guevara

Yiallouros

Kappelhoff

et al. 2010

Journal of Biological Chemistry

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Proteolysis is regulated by inactive (latent) zymogens, with a prosegment preventing access of substrates to the activesite cleft of the enzyme. How latency is maintained often depends on the catalytic mechanism of the protease. For example, in several families of the metzincin metallopeptidases, a "cysteine switch" mechanism involves a conserved prosegment motif with a cysteine residue that coordinates the catalytic zinc ion. Another family of metzincins, the astacins, do not possess a cysteine switch, so latency is maintained by other means. We have solved the high resolution crystal structure of proastacin from the European crayfish, Astacus astacus. Its prosegment is the shortest structurally reported for a metallopeptidase, and it has a unique structure. It runs through the active-site cleft in reverse orientation to a genuine substrate. Moreover, a conserved aspartate, projected by a wide loop of the prosegment, coordinates the zinc ion instead of the catalytic solvent molecule found in the mature enzyme. Activation occurs through two-step limited proteolysis and entails major rearrangement of a flexible activation domain, which becomes rigid and creates the base of the substrate-binding cleft. Maturation also requires the newly formed N terminus to be precisely trimmed so that it can participate in a buried solvent-mediated hydrogen-bonding network, which includes an invariant active-site residue. We describe a novel mechanism for latency and activation, which shares some common features both with other metallopeptidases and with serine peptidases.The proteolytic activity of most metallopeptidases (MPs) 3 is regulated, and it is only exerted where and when required (1). Such control may occur through modulation of gene expression, compartmentalization, allostery, or inhibition by protein inhibitors. Another regulatory mechanism is zymogenic latency, which is provided by mostly N-terminal prosegments. These block access of substrates to the active-site cleft, and they are removed by limited proteolysis during maturation (2, 3). Such prosegments often fold independently and guide on their part the folding process of the cognate protease domains. They may also act as intramolecular chaperones or inhibitors of the mature enzymes in trans and in intracellular sorting of the zymogen (2). Therefore, the study of the molecular mechanisms by which MPs maintain latency is indispensable to an understanding of their basic mode of action. It also paves the way for the design of inhibitors that mimic the latent state so as to modulate MP activity as part of therapeutic approaches. Detailed three-dimensional structural information can contribute much to this understanding (4). However, among the MPs, only funnelins, lysostaphins, thermolysins, and matrix metalloproteinases (MMPs) have been structurally analyzed for their zymogens. Results reveal that the mature enzyme moieties are already in a competent conformation. Notwithstanding, each group displays a distinct mechanism of latency maintenance (5-11).The metzincins...

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Catalytic Domain Architecture of Metzincin Metalloproteases

Cited by 206 publications

References 44 publications

Structure, function and latency regulation of a bacterial enterotoxin potentially derived from a mammalian adamalysin/ADAM xenolog

Structure, function and latency regulation of a bacterial enterotoxin potentially derived from a mammalian adamalysin/ADAM xenolog

Crystal structure of archaemetzincin amza from Methanopyrus kandleri at 1.5 Å resolution

Proenzyme Structure and Activation of Astacin Metallopeptidase

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