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Proteolytic activity regarding Sarconesiopsis magellanica (Diptera: Calliphoridae) larval excretions and secretions.
Acta Trop. 2013 Dec; 128(3):686-91.AT

Abstract

Sarconesiopsis magellanica (Diptera: Calliphoridae) is a medically important necrophagous fly which is used for establishing the post-mortem interval. Diptera maggots release proteolytic enzymes contained in larval excretion and secretion (ES) products playing a key role in digestion. Special interest in proteolytic enzymes has also been aroused regarding understanding their role in wound healing since they degrade necrotic tissue during larval therapy. This study was thus aimed at identifying and characterising S. magellanica proteolytic enzyme ES products for the first time. These products were obtained from first-, second- and third-instar larvae taken from a previously-established colony. ES proteins were separated by SDS-PAGE and their proteolytic activity was characterised by zymograms and inhibition assays involving BAPNA (Nα-benzoyl-dl-Arg-p-nitroanilide) and SAPNA substrates, using synthetic inhibitors. The protein profile ranged from ∼69kDa to ∼23kDa; several of them coincided with the Lucilia sericata ES protein profile. Serine-protease hydrolysis activity (measured by zymogram) was confirmed when a ∼25kDa band disappeared upon ES incubation with PMSF inhibitor at pH 7.8. Analysis of larval ES proteolytic activity on BAPNA and SAPNA substrates (determined by using TLCK and TPCK specific inhibitors) suggested a greater amount of trypsin-like protease. These results support the need for further experiments aimed at validating S. magellanica use in larval therapy.

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Authors+Show Affiliations

Pinilla YT
Medical and Forensic Entomology Research Group, School of Medicine and Health Sciences, Universidad del Rosario, Bogotá, Colombia. Electronic address: tatis_espinita@hotmail.com.
Moreno-Pérez DA
No affiliation info available
Patarroyo MA
No affiliation info available
Bello FJ
No affiliation info available

MeSH

AnimalsBenzoylarginine NitroanilideBodily SecretionsChromogenic CompoundsDipteraElectrophoresis, Polyacrylamide GelEnzyme InhibitorsHumansHydrogen-Ion ConcentrationLarvaMolecular WeightPeptide HydrolasesProteolysis

Pub Type(s)

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

Language

eng

PubMed ID

24076089

Citation

Pinilla, Yudi T., et al. "Proteolytic Activity Regarding Sarconesiopsis Magellanica (Diptera: Calliphoridae) Larval Excretions and Secretions." Acta Tropica, vol. 128, no. 3, 2013, pp. 686-91.
Pinilla YT, Moreno-Pérez DA, Patarroyo MA, et al. Proteolytic activity regarding Sarconesiopsis magellanica (Diptera: Calliphoridae) larval excretions and secretions. Acta Trop. 2013;128(3):686-91.
Pinilla, Y. T., Moreno-Pérez, D. A., Patarroyo, M. A., & Bello, F. J. (2013). Proteolytic activity regarding Sarconesiopsis magellanica (Diptera: Calliphoridae) larval excretions and secretions. Acta Tropica, 128(3), 686-91. https://doi.org/10.1016/j.actatropica.2013.09.020
Pinilla YT, et al. Proteolytic Activity Regarding Sarconesiopsis Magellanica (Diptera: Calliphoridae) Larval Excretions and Secretions. Acta Trop. 2013;128(3):686-91. PubMed PMID: 24076089.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Proteolytic activity regarding Sarconesiopsis magellanica (Diptera: Calliphoridae) larval excretions and secretions. AU - Pinilla,Yudi T, AU - Moreno-Pérez,Darwin A, AU - Patarroyo,Manuel A, AU - Bello,Felio J, Y1 - 2013/09/25/ PY - 2013/05/23/received PY - 2013/09/06/revised PY - 2013/09/18/accepted PY - 2013/10/1/entrez PY - 2013/10/1/pubmed PY - 2014/6/17/medline KW - Larvae KW - Larval therapy KW - Necrotic wounds KW - Proteases KW - Proteolytic activity KW - S. magellanica SP - 686 EP - 91 JF - Acta tropica JO - Acta Trop VL - 128 IS - 3 N2 - Sarconesiopsis magellanica (Diptera: Calliphoridae) is a medically important necrophagous fly which is used for establishing the post-mortem interval. Diptera maggots release proteolytic enzymes contained in larval excretion and secretion (ES) products playing a key role in digestion. Special interest in proteolytic enzymes has also been aroused regarding understanding their role in wound healing since they degrade necrotic tissue during larval therapy. This study was thus aimed at identifying and characterising S. magellanica proteolytic enzyme ES products for the first time. These products were obtained from first-, second- and third-instar larvae taken from a previously-established colony. ES proteins were separated by SDS-PAGE and their proteolytic activity was characterised by zymograms and inhibition assays involving BAPNA (Nα-benzoyl-dl-Arg-p-nitroanilide) and SAPNA substrates, using synthetic inhibitors. The protein profile ranged from ∼69kDa to ∼23kDa; several of them coincided with the Lucilia sericata ES protein profile. Serine-protease hydrolysis activity (measured by zymogram) was confirmed when a ∼25kDa band disappeared upon ES incubation with PMSF inhibitor at pH 7.8. Analysis of larval ES proteolytic activity on BAPNA and SAPNA substrates (determined by using TLCK and TPCK specific inhibitors) suggested a greater amount of trypsin-like protease. These results support the need for further experiments aimed at validating S. magellanica use in larval therapy. SN - 1873-6254 UR - https://www.unboundmedicine.com/medline/citation/24076089/Proteolytic_activity_regarding_Sarconesiopsis_magellanica__Diptera:_Calliphoridae__larval_excretions_and_secretions_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0001-706X(13)00270-2 DB - PRIME DP - Unbound Medicine ER -