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A single amino acid substitution affects substrate specificity in cysteine proteinases from Fasciola hepatica.
Protein Sci. 2000 Dec; 9(12):2567-72.PS

Abstract

The trematode Fasciola hepatica secretes a number of cathepsin L-like proteases that are proposed to be involved in feeding, migration, and immune evasion by the parasite. To date, six full cDNA sequences encoding cathepsin L preproproteins have been identified. Previous studies have demonstrated that one of these cathepsins (L2) is unusual in that it is able to cleave substrates with a proline in the P2 position, translating into an unusual ability (for a cysteine proteinase) to clot fibrinogen. In this study, we report the sequence of a novel cathepsin (L5) and compare the substrate specificity of a recombinant enzyme with that of recombinant cathepsin L2. Despite sharing 80% sequence identity with cathepsin L2, cathepsin L5 does not exhibit substantial catalytic activity against substrates containing proline in the P2 position. Molecular modeling studies suggested that a single amino acid change (L69Y) in the mature proteinases may account for the difference in specificity at the S2 subsite. Recombinant cathepsin L5/L69Y was expressed in yeast and a substantial increase in the ability of this variant to accommodate substrates with a proline residue in the P2 position was observed. Thus, we have identified a single amino acid substitution that can substantially influence the architecture of the S2 subsite of F. hepatica cathepsin L proteases.

Authors+Show Affiliations

Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

11206078

Citation

Smooker, P M., et al. "A Single Amino Acid Substitution Affects Substrate Specificity in Cysteine Proteinases From Fasciola Hepatica." Protein Science : a Publication of the Protein Society, vol. 9, no. 12, 2000, pp. 2567-72.
Smooker PM, Whisstock JC, Irving JA, et al. A single amino acid substitution affects substrate specificity in cysteine proteinases from Fasciola hepatica. Protein Sci. 2000;9(12):2567-72.
Smooker, P. M., Whisstock, J. C., Irving, J. A., Siyaguna, S., Spithill, T. W., & Pike, R. N. (2000). A single amino acid substitution affects substrate specificity in cysteine proteinases from Fasciola hepatica. Protein Science : a Publication of the Protein Society, 9(12), 2567-72.
Smooker PM, et al. A Single Amino Acid Substitution Affects Substrate Specificity in Cysteine Proteinases From Fasciola Hepatica. Protein Sci. 2000;9(12):2567-72. PubMed PMID: 11206078.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A single amino acid substitution affects substrate specificity in cysteine proteinases from Fasciola hepatica. AU - Smooker,P M, AU - Whisstock,J C, AU - Irving,J A, AU - Siyaguna,S, AU - Spithill,T W, AU - Pike,R N, PY - 2001/2/24/pubmed PY - 2001/6/2/medline PY - 2001/2/24/entrez SP - 2567 EP - 72 JF - Protein science : a publication of the Protein Society JO - Protein Sci VL - 9 IS - 12 N2 - The trematode Fasciola hepatica secretes a number of cathepsin L-like proteases that are proposed to be involved in feeding, migration, and immune evasion by the parasite. To date, six full cDNA sequences encoding cathepsin L preproproteins have been identified. Previous studies have demonstrated that one of these cathepsins (L2) is unusual in that it is able to cleave substrates with a proline in the P2 position, translating into an unusual ability (for a cysteine proteinase) to clot fibrinogen. In this study, we report the sequence of a novel cathepsin (L5) and compare the substrate specificity of a recombinant enzyme with that of recombinant cathepsin L2. Despite sharing 80% sequence identity with cathepsin L2, cathepsin L5 does not exhibit substantial catalytic activity against substrates containing proline in the P2 position. Molecular modeling studies suggested that a single amino acid change (L69Y) in the mature proteinases may account for the difference in specificity at the S2 subsite. Recombinant cathepsin L5/L69Y was expressed in yeast and a substantial increase in the ability of this variant to accommodate substrates with a proline residue in the P2 position was observed. Thus, we have identified a single amino acid substitution that can substantially influence the architecture of the S2 subsite of F. hepatica cathepsin L proteases. SN - 0961-8368 UR - https://www.unboundmedicine.com/medline/citation/11206078/A_single_amino_acid_substitution_affects_substrate_specificity_in_cysteine_proteinases_from_Fasciola_hepatica_ DB - PRIME DP - Unbound Medicine ER -