Two Clip Domain Family of Serine Proteases and Serine Protease Homolog from the Fall Webworm, Hyphantria cunea; cDNA Clonning and Characterization

• Published in: Entomological research pp. 253 - 260
• Main Authors: 박두상, 박호용
• Format: Journal Article
• Language: English
• Published: 한국곤충학회 01.12.2004
• Subjects: 생물학
• ISSN: 1748-5967; 1748-5967

Two types of serine proteases and a serine protease homologue cDNAs wereisolated from Hyphantria cunealarvae induced immune response due to an injection of amicroorganism through RT-PCR and cDNA library screening, and their characteristics wereexamined. The isolated cDNAs are composed 2.1 kb, 2.2 kb, and 2.5 kb nucleotide each, whichencoded 388, 390, 580 amino acid residues, and were designated as HcPE-1, HcPE-2 and HcPE-3, respectively. They were revealed as serine proteases or a serine protease homologue with theclip domain through a database search. The deduced amino acid sequence comparison showedhigh homology of 72-78% among them. Six Cys residues of the N-terminal clip domain formingthe disulfide bond, Cys residues of the catalytic domain, and Cys residues forming inter-bridgebetween clip domain and catalytic domain were also well preserved. Three amino acid residues,His, Asp, and Ser, within the active site were perfectly conserved in HcPE-2 and HcPE-3,however, His was replaced with Gln178 in HcPE-1. The Arg residues (HcPE-1, Arg132; HcPE-2,Arg134; HcPE-3, Arg325) known as the activation sites by proteolytic cleavage were preserved wellin all three types of protein. In case of HcPE-3, three continuous clip-like domains existed in theN terminal. As the result of phylogenetic analysis, three clip domain family of protein from H.cunea make groups with arthropod proclotting enzyme precursor. Northern blot analysis showedall three genes were induced through an injection of Escherichia coli, but expression patterns were varied. KCI Citation Count: 0