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Identification and partial characterization of excretory/secretory products with proteolytic activity in Giardia intestinalis.
J Parasitol. 2000 Aug; 86(4):859-62.JP

Abstract

This report examines the presence of proteolytic activity detected in media collected from in vitro cultures of Giardia intestinalis, and the partial characterization by gelatin-substrate polyacrylamide gel electrophoresis and inhibition studies. Gelatin-substrate polyacrylamide gel electrophoresis revealed 6 bands with proteolytic activity, with estimated molecular weights of 36, 59, 63, 72, 103, and 175 kDa. These bands were not present in the control medium. On the other hand, G. intestinalis trophozoite lysates showed proteolytic bands at 16, 20, 66, 82, 108, and 120 kDa, thus indicating that intracellular proteases could be different from the excretory/secretory (E/S) products. Based on inhibition studies, 2 bands of 59 and 63 kDa were inhibited by iodoacetic acid, indicating the presence of cysteine proteases. Partial inhibition of a band of 36 kDa was found with EDTA, a metal-chelating agent, suggesting the possible presence of metalloproteases. The presence of aspartic and serine proteases were not detected under the assay conditions used. As G. intestinalis E/S may be involved in differentiation mechanisms of the parasite and also be responsible for the mucosal alterations that occur in giardiasis, the characterization of these proteases may facilitate their evaluation as targets in the therapy of the disease.

Authors+Show Affiliations

Jiménez JC
Secciones de Immunopatología, Instituto de Biomedicina, Universidad Central de Venezuela, Caracas.
Uzcanga G
No affiliation info available
Zambrano A
No affiliation info available
Di Prisco MC
No affiliation info available
Lynch NR
No affiliation info available

MeSH

AnimalsCulture MediaEdetic AcidElectrophoresis, Polyacrylamide GelGiardia lambliaIodoacetic AcidMolecular WeightPepstatinsPeptide HydrolasesPhenylmethylsulfonyl FluorideProtease Inhibitors

Pub Type(s)

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

Language

eng

PubMed ID

10958474

Citation

Jiménez, J C., et al. "Identification and Partial Characterization of Excretory/secretory Products With Proteolytic Activity in Giardia Intestinalis." The Journal of Parasitology, vol. 86, no. 4, 2000, pp. 859-62.
Jiménez JC, Uzcanga G, Zambrano A, et al. Identification and partial characterization of excretory/secretory products with proteolytic activity in Giardia intestinalis. J Parasitol. 2000;86(4):859-62.
Jiménez, J. C., Uzcanga, G., Zambrano, A., Di Prisco, M. C., & Lynch, N. R. (2000). Identification and partial characterization of excretory/secretory products with proteolytic activity in Giardia intestinalis. The Journal of Parasitology, 86(4), 859-62.
Jiménez JC, et al. Identification and Partial Characterization of Excretory/secretory Products With Proteolytic Activity in Giardia Intestinalis. J Parasitol. 2000;86(4):859-62. PubMed PMID: 10958474.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Identification and partial characterization of excretory/secretory products with proteolytic activity in Giardia intestinalis. AU - Jiménez,J C, AU - Uzcanga,G, AU - Zambrano,A, AU - Di Prisco,M C, AU - Lynch,N R, PY - 2000/8/25/pubmed PY - 2000/9/9/medline PY - 2000/8/25/entrez SP - 859 EP - 62 JF - The Journal of parasitology JO - J Parasitol VL - 86 IS - 4 N2 - This report examines the presence of proteolytic activity detected in media collected from in vitro cultures of Giardia intestinalis, and the partial characterization by gelatin-substrate polyacrylamide gel electrophoresis and inhibition studies. Gelatin-substrate polyacrylamide gel electrophoresis revealed 6 bands with proteolytic activity, with estimated molecular weights of 36, 59, 63, 72, 103, and 175 kDa. These bands were not present in the control medium. On the other hand, G. intestinalis trophozoite lysates showed proteolytic bands at 16, 20, 66, 82, 108, and 120 kDa, thus indicating that intracellular proteases could be different from the excretory/secretory (E/S) products. Based on inhibition studies, 2 bands of 59 and 63 kDa were inhibited by iodoacetic acid, indicating the presence of cysteine proteases. Partial inhibition of a band of 36 kDa was found with EDTA, a metal-chelating agent, suggesting the possible presence of metalloproteases. The presence of aspartic and serine proteases were not detected under the assay conditions used. As G. intestinalis E/S may be involved in differentiation mechanisms of the parasite and also be responsible for the mucosal alterations that occur in giardiasis, the characterization of these proteases may facilitate their evaluation as targets in the therapy of the disease. SN - 0022-3395 UR - https://www.unboundmedicine.com/medline/citation/10958474/Identification_and_partial_characterization_of_excretory/secretory_products_with_proteolytic_activity_in_Giardia_intestinalis_ DB - PRIME DP - Unbound Medicine ER -