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The main proteinases in Dermatobia hominis second and third instars larvae are serine-proteinases.
Vet Parasitol. 2007 Apr 30; 145(3-4):326-31.VP

Abstract

We performed a combination of proteinase assay, either in solution or immobilized in sodium dodecyl sulfate-polyacrylamide gel copolymerized with gelatin, to detect and quantify proteinases of Dermatobia hominis second (L2) and third (L3) instar larvae. In the quantitative assay, we examined proteinase activity by hydrolysis of a panel of peptide bonds specific for the main proteinase classes. We verified that the pGlu-Phe-Leu p-nitroanilide substrate was hydrolyzed by crude extracts of L2 (3.0+/-0.2 nmol h(-1)mg of protein(-1)) and L3 (7.7+/-0.1 nmol h(-1)mg of protein(-1)) and that both activities were partially inhibited by trans-epoxysuccinyl-l-leucylamido-(4-guanidino)butane, 15% and 3%, respectively. Also, we demonstrated that the Nalpha-p-Tosyl-l-Arg methyl ester substrate was hydrolyzed by crude extracts of L2 (117+/-24 nmol h(-1)mg of protein(-1)) and L3 (111+/-10 nmol h(-1)mg of protein(-1)), suggesting a predominance of esterase activity in the crude larval preparation. Interestingly, the specific activity of serine-proteinases was totally inhibited by phenylmethylsulphonyl fluoride in the L3 crude extract, while only 10% of this enzyme class activity was inhibited in the L2 crude extract. The results of the qualitative assays with substrate gels suggested that L2 and L3 larvae express serine-proteinases with similar (13 and 22 kDa) and distinct (50 kDa in L2 and 30 kDa in L3) relative molecular masses. These findings contribute to the biochemical characterization of D. hominis L2 and L3 larvae.

Authors+Show Affiliations

Departamento de Bioquímica e Biologia Molecular, Instituto Oswaldo Cruz, FIOCRUZ, Av. Brasil 4365, CEP 21.045-900, Manguinhos, Rio de Janeiro, RJ, Brazil.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

17293049

Citation

Pires, F A., et al. "The Main Proteinases in Dermatobia Hominis Second and Third Instars Larvae Are Serine-proteinases." Veterinary Parasitology, vol. 145, no. 3-4, 2007, pp. 326-31.
Pires FA, Moya-Borja GE, Barreira JD, et al. The main proteinases in Dermatobia hominis second and third instars larvae are serine-proteinases. Vet Parasitol. 2007;145(3-4):326-31.
Pires, F. A., Moya-Borja, G. E., Barreira, J. D., Pinho, R. T., & Alves, C. R. (2007). The main proteinases in Dermatobia hominis second and third instars larvae are serine-proteinases. Veterinary Parasitology, 145(3-4), 326-31.
Pires FA, et al. The Main Proteinases in Dermatobia Hominis Second and Third Instars Larvae Are Serine-proteinases. Vet Parasitol. 2007 Apr 30;145(3-4):326-31. PubMed PMID: 17293049.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The main proteinases in Dermatobia hominis second and third instars larvae are serine-proteinases. AU - Pires,F A, AU - Moya-Borja,G E, AU - Barreira,J D, AU - Pinho,R T, AU - Alves,C R, Y1 - 2007/02/12/ PY - 2006/09/14/received PY - 2006/12/27/revised PY - 2007/01/02/accepted PY - 2007/2/13/pubmed PY - 2007/11/6/medline PY - 2007/2/13/entrez SP - 326 EP - 31 JF - Veterinary parasitology JO - Vet Parasitol VL - 145 IS - 3-4 N2 - We performed a combination of proteinase assay, either in solution or immobilized in sodium dodecyl sulfate-polyacrylamide gel copolymerized with gelatin, to detect and quantify proteinases of Dermatobia hominis second (L2) and third (L3) instar larvae. In the quantitative assay, we examined proteinase activity by hydrolysis of a panel of peptide bonds specific for the main proteinase classes. We verified that the pGlu-Phe-Leu p-nitroanilide substrate was hydrolyzed by crude extracts of L2 (3.0+/-0.2 nmol h(-1)mg of protein(-1)) and L3 (7.7+/-0.1 nmol h(-1)mg of protein(-1)) and that both activities were partially inhibited by trans-epoxysuccinyl-l-leucylamido-(4-guanidino)butane, 15% and 3%, respectively. Also, we demonstrated that the Nalpha-p-Tosyl-l-Arg methyl ester substrate was hydrolyzed by crude extracts of L2 (117+/-24 nmol h(-1)mg of protein(-1)) and L3 (111+/-10 nmol h(-1)mg of protein(-1)), suggesting a predominance of esterase activity in the crude larval preparation. Interestingly, the specific activity of serine-proteinases was totally inhibited by phenylmethylsulphonyl fluoride in the L3 crude extract, while only 10% of this enzyme class activity was inhibited in the L2 crude extract. The results of the qualitative assays with substrate gels suggested that L2 and L3 larvae express serine-proteinases with similar (13 and 22 kDa) and distinct (50 kDa in L2 and 30 kDa in L3) relative molecular masses. These findings contribute to the biochemical characterization of D. hominis L2 and L3 larvae. SN - 0304-4017 UR - https://www.unboundmedicine.com/medline/citation/17293049/The_main_proteinases_in_Dermatobia_hominis_second_and_third_instars_larvae_are_serine_proteinases_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0304-4017(07)00005-2 DB - PRIME DP - Unbound Medicine ER -