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Molecular cloning and partial characterization of a trypsin-like protein in salivary glands of Lygus hesperus (Hemiptera: Miridae).
Insect Biochem Mol Biol. 2002 Apr; 32(4):455-64.IB

Abstract

Trypsin-like enzymes from the salivary gland complex (SGC) of Lygus hesperus Knight were partially purified by preparative isoelectric focusing (IEF). Enzyme active against Nalpha-benzoyl-L-arginine-p-nitroanilide (BApNA) focused at approximately pH 10 during IEF. This alkaline fraction gave a single activity band when analyzed with casein zymograms. The serine proteinase inhibitors, phenylmethylsulfonyl fluoride (PMSF) and lima bean trypsin inhibitor, completely inhibited or suppressed the caseinolytic activity in the crude salivary gland extract as well as the IEF-purified sample. Chicken egg white trypsin inhibitor also inhibited the IEF-purified sample but was not effective against a major caseinolytic band in the crude salivary gland extract. These data indicated the presence of serine proteinases in the SGC of L. hesperus. Cloning and sequencing of a trypsin-like precursor cDNA provided additional direct evidence for serine proteinases in L. hesperus. The encoded trypsin-like protein included amino acid sequence motifs, which are conserved with five homologous serine proteinases from other insects. Typical features of the putative trypsin-like protein from L. hesperus included residues in the serine proteinase active site (His(89), Asp(139), Ser(229)), conserved cysteine residues for disulfide bridges, residues (Asp(223), Gly(252), Gly(262)) that determine trypsin specificity, and both zymogen signal and activation peptides.

Authors+Show Affiliations

Biological Control and Mass Rearing Research Unit, USDA/ARS, P.O. Box 5367, Mississippi State, MS 39762, USA. fzeng@bcmrru.msstate.eduNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

11886780

Citation

Zeng, Fanrong, et al. "Molecular Cloning and Partial Characterization of a Trypsin-like Protein in Salivary Glands of Lygus Hesperus (Hemiptera: Miridae)." Insect Biochemistry and Molecular Biology, vol. 32, no. 4, 2002, pp. 455-64.
Zeng F, Zhu YC, Cohen AC. Molecular cloning and partial characterization of a trypsin-like protein in salivary glands of Lygus hesperus (Hemiptera: Miridae). Insect Biochem Mol Biol. 2002;32(4):455-64.
Zeng, F., Zhu, Y. C., & Cohen, A. C. (2002). Molecular cloning and partial characterization of a trypsin-like protein in salivary glands of Lygus hesperus (Hemiptera: Miridae). Insect Biochemistry and Molecular Biology, 32(4), 455-64.
Zeng F, Zhu YC, Cohen AC. Molecular Cloning and Partial Characterization of a Trypsin-like Protein in Salivary Glands of Lygus Hesperus (Hemiptera: Miridae). Insect Biochem Mol Biol. 2002;32(4):455-64. PubMed PMID: 11886780.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular cloning and partial characterization of a trypsin-like protein in salivary glands of Lygus hesperus (Hemiptera: Miridae). AU - Zeng,Fanrong, AU - Zhu,Yu Cheng, AU - Cohen,Allen C, PY - 2002/3/12/pubmed PY - 2002/5/30/medline PY - 2002/3/12/entrez SP - 455 EP - 64 JF - Insect biochemistry and molecular biology JO - Insect Biochem Mol Biol VL - 32 IS - 4 N2 - Trypsin-like enzymes from the salivary gland complex (SGC) of Lygus hesperus Knight were partially purified by preparative isoelectric focusing (IEF). Enzyme active against Nalpha-benzoyl-L-arginine-p-nitroanilide (BApNA) focused at approximately pH 10 during IEF. This alkaline fraction gave a single activity band when analyzed with casein zymograms. The serine proteinase inhibitors, phenylmethylsulfonyl fluoride (PMSF) and lima bean trypsin inhibitor, completely inhibited or suppressed the caseinolytic activity in the crude salivary gland extract as well as the IEF-purified sample. Chicken egg white trypsin inhibitor also inhibited the IEF-purified sample but was not effective against a major caseinolytic band in the crude salivary gland extract. These data indicated the presence of serine proteinases in the SGC of L. hesperus. Cloning and sequencing of a trypsin-like precursor cDNA provided additional direct evidence for serine proteinases in L. hesperus. The encoded trypsin-like protein included amino acid sequence motifs, which are conserved with five homologous serine proteinases from other insects. Typical features of the putative trypsin-like protein from L. hesperus included residues in the serine proteinase active site (His(89), Asp(139), Ser(229)), conserved cysteine residues for disulfide bridges, residues (Asp(223), Gly(252), Gly(262)) that determine trypsin specificity, and both zymogen signal and activation peptides. SN - 0965-1748 UR - https://www.unboundmedicine.com/medline/citation/11886780/Molecular_cloning_and_partial_characterization_of_a_trypsin_like_protein_in_salivary_glands_of_Lygus_hesperus__Hemiptera:_Miridae__ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0965174801001230 DB - PRIME DP - Unbound Medicine ER -