Characterization of the molecular features and expression patterns of two serine proteases in Hermetia illucens (Diptera: Stratiomyidae) larvae
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To investigate the molecular scavenging capabilities of the larvae of Hermetia illucens, two serine proteases (SPs) were cloned and characterized. Multiple sequence alignments and phylogenetic tree analysis of the deduced amino acid sequences of Hi-SP1 and Hi-SP2 were suggested that Hi-SP1 may be a chymotrypsin- and Hi-SP2 may be a trypsin-like protease. Hi-SP1 and Hi-SP2 3-D homology models revealed that a catalytic triad, three disulfide bonds, and a substrate-binding pocket were highly conserved, as would be expected of a SP. E. coli expressed Hi-SP1 and Hi-SP2 showed chymotrypsin or trypsin activities, respectively. Hi-SP2 mRNAs were consistently expressed during larval development. In contrast, the expression of Hi-SP1 mRNA fluctuated between feeding and molting stages and disappeared at the pupal stages. These expression pattern differences suggest that Hi-SP1 may be a larval specific chymotrypsin-like protease involved with food digestion,while Hi-SP2 may be a trypsin-like protease with diverse functions at different stages. [BMB reports 2011; 44(6):387-392]
Abstract
To investigate the molecular scavenging capabilities of the larvae of Hermetia illucens, two serine proteases (SPs) were cloned and characterized. Multiple sequence alignments and phylogenetic tree analysis of the deduced amino acid sequences of Hi-SP1 and Hi-SP2 were suggested that Hi-SP1 may be a chymotrypsin- and Hi-SP2 may be a trypsin-like protease. Hi-SP1 and Hi-SP2 3-D homology models revealed that a catalytic triad, three disulfide bonds, and a substrate-binding pocket were highly conserved, as would be expected of a SP. E. coli expressed Hi-SP1 and Hi-SP2 showed chymotrypsin or trypsin activities, respectively. Hi-SP2 mRNAs were consistently expressed during larval development. In contrast, the expression of Hi-SP1 mRNA fluctuated between feeding and molting stages and disappeared at the pupal stages. These expression pattern differences suggest that Hi-SP1 may be a larval specific chymotrypsin-like protease involved with food digestion,while Hi-SP2 may be a trypsin-like protease with diverse functions at different stages. [BMB reports 2011; 44(6):387-392]
참고문헌 (18)
- Yan, J, "Molecular characterization of two serine proteases expressed in gut tissue of the African trypanosome vector, Glossina morsitans morsitans", Insect Mol. Biol, 10, pp.47~56 (2001)
- Zhu, Y. C, "Molecular cloning of trypsin-like cDNAs and comparison of proteinase activities in the salivary glands and gut of the tarnished plant bug Lygus lineolaris (Heteroptera: Miridae)", Insect Biochem. Mol. Biol, 33, pp.889~899 (2003)
- Rawlings, N. D, "Families of serine peptidases", Methods Enzymol, 244, pp.19~61 (1994)
- Noriega, F. G, "A molecular view of trypsin synthesis in the midgut of Aedes aegypti", J. Insect Physiol, 45, pp.613~620 (1999)
- Diener, S, "Conversion of organic material by black soldier fly larvae: establishing optimal feeding rates", Waste Manag. Res, 27, pp.603~610 (2009)
- Schmidt, A, "Trypsin revisited: crystallography AT (SUB) atomic resolution and quantum chemistry revealing details of catalysis", J. Biol. Chem, 278, pp.43357~43362 (2003)
- Krem, M. M, "Molecular markers of serine protease evolution", EMBO J, 20, pp.3036~3045 (2001)
- Benkert, P, "QMEAN: A comprehensive scoring function for model quality assessment", Proteins, 71, pp.261~277 (2008)
- Kraut,J, "Serine proteases: structure and mechanism of catalysis", Ann. Rev. Biochem, 46, pp.331~358 (1977)
- Kang, D, "An azurocidin-like protein is induced in Trichoplusia ni larval gut cells after bacterial challenge", Dev. Comp. Immunol, 26, pp.495~503 (2002)
- Sui, Y. P, "The impacts of classical insect hormones on the expression profiles of a new digestive trypsin-like protease (TLP) from the cotton bollworm, Helicoverpa armigera", Insect Mol. Biol, 18, pp.443~452 (2009)
- Lehane, S. M, "Cloning, sequencing, temporal expression and tissue-specificity of two serine proteases from the midgut of the blood-feeding fly Stomoxys calcitrans", Eur. J. Biochem, 254, pp.290~296 (1998)
- Taylor, M. A, "Trypsin isolated from the midgut of the tobacco hornworm, Manduca sexta, is inhibited by synthetic pro-peptides in vitro", Biochem. Biophys. Res. Commun, 235, pp.606~609 (1997)
- Liu, Y, "Characterization of the trypsin-like protease (Ha-TLP2) constitutively expressed in the integument of the cotton bollworm, Helicoverpa armigera", Arch. Insect. Biochem. Physiol, 72, pp.74~87 (2009)
- "Insect Biochem. Mol. Biol", 2005, 23~32
- Cuc, H. T, "Generation and characterization of monoclonal antibodies specific to Drosophila presenilin", Hybridoma (Larchmt), 28, pp.215~220 (2009)
- Lee, D. W, "A Combination of Biochemical and Proteomic Analyses Reveals Bx-LEC-1 as an Antigenic Target for the Monoclonal Antibody 3-2A7-2H5-D9-F10 Specific to the Pine Wood Nematode", MOLECULAR CELLULAR PROTEOMICS, 10(2) (2011), AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
- Youngho Koh, "Biochemical characterization of digestive enzymes in the black soldier fly, Hermetia illucens (Diptera: Stratiomyidae)", Journal of Asia-Pacific Entomology, 14(1), pp.11~14 (2011), 한국응용곤충학회