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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. 2003 Sep; 33(9):889-99.IB

Abstract

Using specific proteinase inhibitors, we demonstrated that serine proteinases in the tarnished plant bug, Lygus lineolaris, are major proteinases in both salivary glands and gut tissues. Gut proteinases were less sensitive to inhibition than proteinases from the salivary glands. Up to 80% azocaseinase and 90% of BApNAse activities in the salivary glands were inhibited by aprotinin, benzamidine, and PMSF, whereas only 46% azocaseinase and 60% BApNAse activities in the gut were suppressed by benzamidine, leupeptin, and TLCK. The pH optima for azocaseinase activity in salivary glands ranged from 6.2 to 10.6, whereas the pH optima for gut proteinases was acidic for general and alkaline for tryptic proteinases. Zymogram analysis demonstrated that approximately 26-kDa proteinases from salivary glands were active against both gelatin and casein substrates. Three trypsin-like cDNAs, LlSgP2-4, and one trypsin-like cDNA, L1GtP1, were cloned from salivary glands and gut, respectively. Putative trypsin precursors from all cloned cDNAs contained a signal peptide, activation peptide, and conserved N-termini (IVGG). Other structural features included His, Asp, and Ser residues for the catalytic amino acid triad of serine proteinase active sites, residues for the binding pocket, and four pairs of cysteine residues for disulfide bridges. Deduced trypsin-like proteins from LlSgP2, LlSgP3, and LlGtP1 cDNAs shared 98-99% sequence identity with a previously reported trypsin-like precursor, whereas the trypsin-like protein of LlSgP4 shared only 44% sequence identity with all other trypsin-like proteins, indicating multi-trypsin forms are present in L. lineolaris.

Authors+Show Affiliations

USDA-ARS-JWDSRC, PO Box 346, Stoneville, MS 38776, USA. yczhu@ars.usda.govNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

12915180

Citation

Zhu, Yu Cheng, et al. "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 Biochemistry and Molecular Biology, vol. 33, no. 9, 2003, pp. 889-99.
Zhu YC, Zeng F, Oppert B. 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. 2003;33(9):889-99.
Zhu, Y. C., Zeng, F., & Oppert, B. (2003). 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 Biochemistry and Molecular Biology, 33(9), 889-99.
Zhu YC, Zeng F, Oppert B. 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. 2003;33(9):889-99. PubMed PMID: 12915180.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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). AU - Zhu,Yu Cheng, AU - Zeng,Fanrong, AU - Oppert,Brenda, PY - 2003/8/14/pubmed PY - 2003/12/3/medline PY - 2003/8/14/entrez SP - 889 EP - 99 JF - Insect biochemistry and molecular biology JO - Insect Biochem Mol Biol VL - 33 IS - 9 N2 - Using specific proteinase inhibitors, we demonstrated that serine proteinases in the tarnished plant bug, Lygus lineolaris, are major proteinases in both salivary glands and gut tissues. Gut proteinases were less sensitive to inhibition than proteinases from the salivary glands. Up to 80% azocaseinase and 90% of BApNAse activities in the salivary glands were inhibited by aprotinin, benzamidine, and PMSF, whereas only 46% azocaseinase and 60% BApNAse activities in the gut were suppressed by benzamidine, leupeptin, and TLCK. The pH optima for azocaseinase activity in salivary glands ranged from 6.2 to 10.6, whereas the pH optima for gut proteinases was acidic for general and alkaline for tryptic proteinases. Zymogram analysis demonstrated that approximately 26-kDa proteinases from salivary glands were active against both gelatin and casein substrates. Three trypsin-like cDNAs, LlSgP2-4, and one trypsin-like cDNA, L1GtP1, were cloned from salivary glands and gut, respectively. Putative trypsin precursors from all cloned cDNAs contained a signal peptide, activation peptide, and conserved N-termini (IVGG). Other structural features included His, Asp, and Ser residues for the catalytic amino acid triad of serine proteinase active sites, residues for the binding pocket, and four pairs of cysteine residues for disulfide bridges. Deduced trypsin-like proteins from LlSgP2, LlSgP3, and LlGtP1 cDNAs shared 98-99% sequence identity with a previously reported trypsin-like precursor, whereas the trypsin-like protein of LlSgP4 shared only 44% sequence identity with all other trypsin-like proteins, indicating multi-trypsin forms are present in L. lineolaris. SN - 0965-1748 UR - https://www.unboundmedicine.com/medline/citation/12915180/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__ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0965174803000948 DB - PRIME DP - Unbound Medicine ER -