Yandamuri_2014_Insects_5_762

Reference

Title : Cloning, Expression, Sequence Analysis and Homology Modeling of the Prolyl Endoprotease from Eurygaster integriceps Puton - Yandamuri_2014_Insects_5_762
Author(s) : Yandamuri RC , Gautam R , Darkoh C , Dareddy V , El-Bouhssini M , Clack BA
Ref : Insects , 5 :762 , 2014
Abstract :

eurygaster integriceps Puton, commonly known as sunn pest, is a major pest of wheat in Northern Africa, the Middle East and Eastern Europe. This insect injects a prolyl endoprotease into the wheat, destroying the gluten. The purpose of this study was to clone the full length cDNA of the sunn pest prolyl endoprotease (spPEP) for expression in E. coli and to compare the amino acid sequence of the enzyme to other known PEPs in both phylogeny and potential tertiary structure. Sequence analysis shows that the 5' UTR contains several putative transcription factor binding sites for transcription factors known to be expressed in Drosophila that might be useful targets for inhibition of the enzyme. The spPEP was first identified as a prolyl endoprotease by Darkoh et al., 2010. The enzyme is a unique serine protease of the S9A family by way of its substrate recognition of the gluten proteins, which are greater than 30 kD in size. At 51% maximum identity to known PEPs, homology modeling using SWISS-MODEL, the porcine brain PEP (PDB: 2XWD) was selected in the database of known PEP structures, resulting in a predicted tertiary structure 99% identical to the porcine brain PEP structure. A Km for the recombinant spPEP was determined to be 210 +/- 53 microM for the zGly-Pro-pNA substrate in 0.025 M ethanolamine, pH 8.5, containing 0.1 M NaCl at 37 C with a turnover rate of 172 +/- 47 microM Gly-Pro-pNA/s/microM of enzyme.

PubMedSearch : Yandamuri_2014_Insects_5_762
PubMedID: 26462938
Gene_locus related to this paper: 9hemi-e1u339

Related information

Substrate Z-Gly-Pro-pNA
Gene_locus 9hemi-e1u339

Citations formats

Yandamuri RC, Gautam R, Darkoh C, Dareddy V, El-Bouhssini M, Clack BA (2014)
Cloning, Expression, Sequence Analysis and Homology Modeling of the Prolyl Endoprotease from Eurygaster integriceps Puton
Insects 5 :762

Yandamuri RC, Gautam R, Darkoh C, Dareddy V, El-Bouhssini M, Clack BA (2014)
Insects 5 :762