PDBsum entry: 2o6x

Hydrolase

PDB id: 2o6x

Contents

Protein chain

306 a.a. *

Waters ×388

* Residue conservation analysis

PDB id:

2o6x

Name:

Hydrolase

Title:

Crystal structure of procathepsin l1 from fasciola hepatica

Structure:

Secreted cathepsin l 1. Chain: a. Synonym: procathepsin l1. Engineered: yes. Mutation: yes. Other_details: inactive zymogen form

Source:

Fasciola hepatica. Liver fluke. Organism_taxid: 6192. Strain: irish isolate. Gene: cl1. Expressed in: pichia pastoris. Expression_system_taxid: 4922.

Resolution:

1.40Å R-factor: 0.129 R-free: 0.165

Authors:

L.S.Brinen,J.P.Dalton,S.Geiger,R.Marion,C.M.Stack

Key ref:

C.M.Stack et al. (2008). Structural and functional relationships in the virulence-associated cathepsin L proteases of the parasitic liver fluke, Fasciola hepatica. J Biol Chem, 283, 9896-9908. PubMed id: 18160404 DOI: 10.1074/jbc.M708521200

Date:

08-Dec-06 Release date: 18-Dec-07

Protein chain

Q24940  (CATLL_FASHE) -  Cathepsin L-like proteinase

Seq: 326 a.a.

Struc: 306 a.a.*

* PDB and UniProt seqs differ at 9 residue positions (black crosses)

 Gene Ontology (GO) functional annotation 

• Cellular component: extracellular region (1 term)
• Biological process: proteolysis (1 term)
• Biochemical function: hydrolase activity (5 terms)

DOI no: 10.1074/jbc.M708521200

J Biol Chem 283:9896-9908 (2008)

PubMed id: 18160404

Structural and functional relationships in the virulence-associated cathepsin L proteases of the parasitic liver fluke, Fasciola hepatica.

C.M.Stack, C.R.Caffrey, S.M.Donnelly, A.Seshaadri, J.Lowther, J.F.Tort, P.R.Collins, M.W.Robinson, W.Xu, J.H.McKerrow, C.S.Craik, S.R.Geiger, R.Marion, L.S.Brinen, J.P.Dalton.

ABSTRACT

The helminth parasite Fasciola hepatica secretes cysteine proteases to facilitate tissue invasion, migration, and development within the mammalian host. The major proteases cathepsin L1 (FheCL1) and cathepsin L2 (FheCL2) were recombinantly produced and biochemically characterized. By using site-directed mutagenesis, we show that residues at position 67 and 205, which lie within the S2 pocket of the active site, are critical in determining the substrate and inhibitor specificity. FheCL1 exhibits a broader specificity and a higher substrate turnover rate compared with FheCL2. However, FheCL2 can efficiently cleave substrates with a Pro in the P2 position and degrade collagen within the triple helices at physiological pH, an activity that among cysteine proteases has only been reported for human cathepsin K. The 1.4-A three-dimensional structure of the FheCL1 was determined by x-ray crystallography, and the three-dimensional structure of FheCL2 was constructed via homology-based modeling. Analysis and comparison of these structures and our biochemical data with those of human cathepsins L and K provided an interpretation of the substrate-recognition mechanisms of these major parasite proteases. Furthermore, our studies suggest that a configuration involving residue 67 and the "gatekeeper" residues 157 and 158 situated at the entrance of the active site pocket create a topology that endows FheCL2 with its unusual collagenolytic activity. The emergence of a specialized collagenolytic function in Fasciola likely contributes to the success of this tissue-invasive parasite.

Selected figure(s)

Figure 6.
FIGURE 6. A, bi-lobed mature FheproCL1 Gly^25 is shown as a schematic. The predominantly helical domain is at left, and the predominantly sheet domain is at right. The mutated active site residue Gly^25 lies in the cleft between the two domains and is indicated in red. B, structure of full-length FheproCL1 Gly^25 zymogen is shown, with the mature segment surface illustrated in blue and the prosegment as a schematic. The extended C-terminal portion of the prosegment runs through the active site cleft. The catalytic machinery of the enzyme is highlighted in pink. Figures were created with PyMol (42).

Figure 8.
FIGURE 8. Comparison of the substrate specificity of human cathepsin L (CL), human cathepsin K (CK), recombinant FheCL1, FheCL1 L205A (L205A), FheCL1 L67Y (L67Y), and FheCL2. Data shown as relative k[cat]/K[m] for the hydrolysis of the substrates Z-Phe-Arg-NHMec, Z-Phe-Leu-NHMec, and Tos-Gly-Pro-ArgNHMec. Asterisk indicates data for human CL and CK are taken from Lecaille et al. (44).

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2008, 283, 9896-9908) copyright 2008.

Figures were selected by an automated process.