Tags

Type your tag names separated by a space and hit enter

Midgut proteases from Mamestra configurata (Lepidoptera: Noctuidae) larvae: characterization, cDNA cloning, and expressed sequence tag analysis.
Arch Insect Biochem Physiol. 2003 May; 53(1):30-47.AI

Abstract

The activities of digestive protease within the midgut of Mamestra configurata (bertha armyworm) larvae were examined using specific substrates and protease inhibitors. The bulk of the activity was associated with serine proteases comprising trypsin-, chymotrypsin-, and elastase-like enzymes. At least 10-15 serine protease isozymes were detected using one-dimension gelatin gel electrophoresis. Cysteine or aspartic protease activities were not present; however, amino- and carboxypeptidase activities were associated with the midgut extract. Midgut proteases were active in the pH range of 5.0-12.0 with peaks at pH 7.5 and 11.0. In general, the middle region of the midgut exhibited a higher pH (approximately 8.0) than either the posterior or anterior regions (approximately 7.3-7.7). Moulting larvae possessed a neutral gut pH that was 0.5-1.5 units below that of feeding larvae. Degenerate PCR and expressed sequence tag (EST)-based approaches were used to isolate 30 distinct serine protease encoding cDNAs from a midgut-specific cDNA library including 8 putative trypsins, 9 chymotrypsins, 1 elastase, and 12 whose potential activities could not be determined. cDNAs encoding three amino- and two carboxypeptidases were also identified. Larvae feeding upon artificial diet containing 0.2% soybean trypsin inhibitor experienced a significant delay in development.

Authors+Show Affiliations

Agriculture and Agri-Food Canada, Saskatoon, Canada. hegedusd@em.agr.caNo affiliation info availableNo affiliation info availableNo affiliation info availableNo 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

12701112

Citation

Hegedus, Dwayne, et al. "Midgut Proteases From Mamestra Configurata (Lepidoptera: Noctuidae) Larvae: Characterization, cDNA Cloning, and Expressed Sequence Tag Analysis." Archives of Insect Biochemistry and Physiology, vol. 53, no. 1, 2003, pp. 30-47.
Hegedus D, Baldwin D, O'Grady M, et al. Midgut proteases from Mamestra configurata (Lepidoptera: Noctuidae) larvae: characterization, cDNA cloning, and expressed sequence tag analysis. Arch Insect Biochem Physiol. 2003;53(1):30-47.
Hegedus, D., Baldwin, D., O'Grady, M., Braun, L., Gleddie, S., Sharpe, A., Lydiate, D., & Erlandson, M. (2003). Midgut proteases from Mamestra configurata (Lepidoptera: Noctuidae) larvae: characterization, cDNA cloning, and expressed sequence tag analysis. Archives of Insect Biochemistry and Physiology, 53(1), 30-47.
Hegedus D, et al. Midgut Proteases From Mamestra Configurata (Lepidoptera: Noctuidae) Larvae: Characterization, cDNA Cloning, and Expressed Sequence Tag Analysis. Arch Insect Biochem Physiol. 2003;53(1):30-47. PubMed PMID: 12701112.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Midgut proteases from Mamestra configurata (Lepidoptera: Noctuidae) larvae: characterization, cDNA cloning, and expressed sequence tag analysis. AU - Hegedus,Dwayne, AU - Baldwin,Doug, AU - O'Grady,Michael, AU - Braun,Lorraine, AU - Gleddie,Steve, AU - Sharpe,Andrew, AU - Lydiate,Derek, AU - Erlandson,Martin, PY - 2003/4/18/pubmed PY - 2003/6/5/medline PY - 2003/4/18/entrez SP - 30 EP - 47 JF - Archives of insect biochemistry and physiology JO - Arch Insect Biochem Physiol VL - 53 IS - 1 N2 - The activities of digestive protease within the midgut of Mamestra configurata (bertha armyworm) larvae were examined using specific substrates and protease inhibitors. The bulk of the activity was associated with serine proteases comprising trypsin-, chymotrypsin-, and elastase-like enzymes. At least 10-15 serine protease isozymes were detected using one-dimension gelatin gel electrophoresis. Cysteine or aspartic protease activities were not present; however, amino- and carboxypeptidase activities were associated with the midgut extract. Midgut proteases were active in the pH range of 5.0-12.0 with peaks at pH 7.5 and 11.0. In general, the middle region of the midgut exhibited a higher pH (approximately 8.0) than either the posterior or anterior regions (approximately 7.3-7.7). Moulting larvae possessed a neutral gut pH that was 0.5-1.5 units below that of feeding larvae. Degenerate PCR and expressed sequence tag (EST)-based approaches were used to isolate 30 distinct serine protease encoding cDNAs from a midgut-specific cDNA library including 8 putative trypsins, 9 chymotrypsins, 1 elastase, and 12 whose potential activities could not be determined. cDNAs encoding three amino- and two carboxypeptidases were also identified. Larvae feeding upon artificial diet containing 0.2% soybean trypsin inhibitor experienced a significant delay in development. SN - 0739-4462 UR - https://www.unboundmedicine.com/medline/citation/12701112/Midgut_proteases_from_Mamestra_configurata__Lepidoptera:_Noctuidae__larvae:_characterization_cDNA_cloning_and_expressed_sequence_tag_analysis_ L2 - https://doi.org/10.1002/arch.10084 DB - PRIME DP - Unbound Medicine ER -